Impact of Liver Inflammation on Bile Acid Side Chain Shortening and Amidation

Bile acid; Inflammation; Oncostatin MÀcid biliar; Inflamació; Oncostatina MÁcido biliar; Inflamación; Oncostatina MBile acid (BA) synthesis from cholesterol by hepatocytes is inhibited by inflammatory cytokines. Whether liver inflammation also affects BA side chain shortening and conjugation was investigated. In human liver cell lines (IHH, HepG2, and HepaRG), agonists of nuclear receptors including the farnesoid X receptor (FXR), liver X receptor (LXR), and peroxisome proliferator-activated receptors (PPARs) did not affect the expression of BA-related peroxisomal enzymes. In contrast, hepatocyte nuclear factor 4α (HNF4α) inhibition down-regulated acyl-CoA oxidase 2 (ACOX2). ACOX2 was repressed by fibroblast growth factor 19 (FGF19), which was prevented by extracellular signal-regulated kinase (ERK) pathway inhibition. These changes were paralleled by altered BA synthesis (HPLC-MS/MS). Cytokines able to down-regulate cholesterol-7α-hydroxylase (CYP7A1) had little effect on peroxisomal enzymes involved in BA synthesis except for ACOX2 and bile acid-CoA:amino acid N-acyltransferase (BAAT), which were down-regulated, mainly by oncostatin M (OSM). This effect was prevented by Janus kinase (JAK) inhibition, which restored BA side chain shortening and conjugation. The binding of OSM to the extracellular matrix accounted for a persistent effect after culture medium replacement. In silico analysis of four databases (n = 201) and a validation cohort (n = 90) revealed an inverse relationship between liver inflammation and ACOX2/BAAT expression which was associated with changes in HNF4α levels. In conclusion, BA side chain shortening and conjugation are inhibited by inflammatory effectors. However, other mechanisms involved in BA homeostasis counterbalance any significant impact on the serum BA profile.This study was supported by the CIBERehd (EHD15PI05/2016) and Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III, Spain (PI19/00819, PI20/00189, and PI20/01663 co-funded by European Regional Development Fund/European Social Fund, “Investing in your future”); Junta de Castilla y Leon (SA074P20); Fundació Marato TV3 (Ref. 201916/31), Spain; AECC Scientific Foundation (2017/2020), Spain; Interdisciplinary Center for Clinical Research (IZKF) at the University Hospital of Wuerzburg, Germany (Project A-E-384 to H.M.H.); grants PID2019-111669-RB-I00, PID2020-115055RB-I00 from Agencia Estatal de Investigación (AEI), Spain; the AGAUR of the Generalidad de Cataluña SGR-2017-1112, Spain; and European Cooperation in Science & Technology (COST) Action CA17112. R.E.E was recipient of a predoctoral fellowship from “Junta de Castilla y León” and “Fondo Social Europeo” (EDU/574/2018). J.A. was recipient of a grant from Fundación Echebano (2020–2022)

An Evaluation of Landmark-Based Methods to Explore Tooth Score Morphology: A Case Study on Felids and Hyenids

Taphonomic studies aim to identify the modifying agents that intervene in bone assemblages found at archaeopaleontological sites. Carnivores may modify, accumulate, or scavenge skeletal parts inflicting tooth marks, including scores, on the cortical surface. Several works have studied tooth score morphology to discern which carnivore group modified the bone assemblages, achieving different results. In the present study, different methods based on the use of landmarks and semilandmarks have been tested to describe and analyze the score profile cross-sections of spotted and brown hyenas, leopards, and lions. According to our results, the already published seven-landmark method is useful in order to differentiate between carnivore species from different families (e.g., felids and hyenids). Meanwhile, felid species (e.g., leopards and lions) cannot be consistently distinguished using any of the methods tested here. In contrast, hyenid species can be morphologically differentiated. On the other hand, the use of semilandmarks does not generally improve morphological characterization and distinction, but low numbers of landmarks and the inclusion of the score’s deepest point might provide the best results when semi-automatic semilandmark models are preferred to avoid sampling biases.The grant IJC2020-043576-I funded by MCIN/AEI/10.13039/501100011033 and the “European Union NextGenerationEU/PRTR” has been awarded to M.C.A. The grant RYC2021-034813-I funded by MCIN/AEI/10.13039/501100011033 and the European Union “NextGenerationEU”/PRTR has been awarded to M.Á.M.-G. During the development of the present work, J.A. was funded by the Euskal Herriko Unibertsitatea [ESPDOC21/05]. L.A.C. is funded by the Spanish Ministry of Science, Innovation and Universities by an FPI Predoctoral grant PRE2019-089411 associated with project RTI2018-099850-B-I0

A remarkable synergistic effect at the transcriptomic level in peach fruits doubly infected by Prunus necrotic ringspot virus and Peach latent mosaic viroid

[EN] Background: Microarray profiling is a powerful technique to investigate expression changes of large amounts of genes in response to specific environmental conditions. The majority of the studies investigating gene expression changes in virus-infected plants are limited to interactions between a virus and a model host plant, which usually is Arabidopsis thaliana or Nicotiana benthamiana. In the present work, we performed microarray profiling to explore changes in the expression profile of field-grown Prunus persica (peach) originating from Chile upon single and double infection with Prunus necrotic ringspot virus (PNRSV) and Peach latent mosaic viroid (PLMVd), worldwide natural pathogens of peach trees. Results: Upon single PLMVd or PNRSV infection, the number of statistically significant gene expression changes was relatively low. By contrast, doubly-infected fruits presented a high number of differentially regulated genes. Among these, down-regulated genes were prevalent. Functional categorization of the gene expression changes upon double PLMVd and PNRSV infection revealed protein modification and degradation as the functional category with the highest percentage of repressed genes whereas induced genes encoded mainly proteins related to phosphate, C-compound and carbohydrate metabolism and also protein modification. Overrepresentation analysis upon double infection with PLMVd and PNRSV revealed specific functional categories over- and underrepresented among the repressed genes indicating active counter-defense mechanisms of the pathogens during infection. Conclusions: Our results identify a novel synergistic effect of PLMVd and PNRSV on the transcriptome of peach fruits. We demonstrate that mixed infections, which occur frequently in field conditions, result in a more complex transcriptional response than that observed in single infections. Thus, our data demonstrate for the first time that the simultaneous infection of a viroid and a plant virus synergistically affect the host transcriptome in infected peach fruits. These field studies can help to fully understand plant-pathogen interactions and to develop appropriate crop protection strategies.We thank Drs M.A. Perez-Amador y J. Gadea for helping in the result analysis. This work was supported by grant BIO2011-25018 from the Spanish granting agency Direccion General de Investigacion Cientifica for the transcriptomic analyses and from the grant 2009CL0020 from the bilateral project INIA-Chile/CSIC-Spain for the phytosanitary evaluation. MC Herranz was the recipient of a contract from the Juan de la Cierva program of the Ministerio de Educacion y Ciencia of Spain.Herranz Gordo, MDC.; Niehl, A.; Rosales, M.; Fiore, N.; Zamorano, A.; Granell Richart, A.; Pallás Benet, V. (2013). A remarkable synergistic effect at the transcriptomic level in peach fruits doubly infected by Prunus necrotic ringspot virus and Peach latent mosaic viroid. Virology Journal. 10:11-15. https://doi.org/10.1186/1743-422X-10-164S111510Pallas V, Garcia JA: How do plant viruses induce disease? Interactions and interference with host components. J Gen Virol 2011, 92: 2691-2705.Whitham SA, Yang C, Goodin MM: Global impact: elucidating plant responses to viral infection. Mol Plant Microbe Interact 2006, 19: 1207-1215.Havelda Z, Varallyay E, Valoczi A, Burgyan J: Plant virus infection-induced persistent host gene downregulation in systemically infected leaves. Plant J 2008, 55: 278-288.Aranda M, Maule A: Virus-induced host gene shutoff in animals and plants. Virology 1998, 243: 261-267.Whitham SA, Quan S, Chang HS, Cooper B, Estes B, Zhu T, Wang X, Hou YM: Diverse RNA viruses elicit the expression of common sets of genes in susceptible Arabidopsis thaliana plants. Plant J 2003, 33: 271-283.Liu Y, Ren D, Pike S, Pallardy S, Gassmann W, Zhang S: Chloroplast-generated reactive oxygen species are involved in hypersensitive response-like cell death mediated by a mitogen-activated protein kinase cascade. Plant J 2007, 51: 941-954.Hadidi A, Barba M: Economic impact of pome and stone fruit viruses and viroids. In Virus and Virus Like Diseases of Pome and Stone Fruits. Edited by: Hadidi A, Barba M, Candresse T, Jelkmann W. St Paul, MN: American Phytopathological Society; 2011:1-8.Flores R, Delgado S, Rodio ME, Ambros S, Hernandez C, Serio FD: Peach latent mosaic viroid: not so latent. Mol Plant Pathol 2006, 7: 209-221.Pallas V, Aparicio F, Herranz MC, Amari K, Sanchez-Pina MA, Myrta A, Sanchez-Navarro JA: Ilarviruses of Prunus spp.: A continued concern for fruit trees. Phytopathology 2012,102(12):1108-1120.Rowland O, Jones JD: Unraveling regulatory networks in plant defense using microarrays. Genome Biol 2001,2(1):1001.1-1001.3.Trinks D, Rajeswaran R, Shivaprasad PV, Akbergenov R, Oakeley EJ, Veluthambi K, Hohn T, Pooggin MM: Suppression of RNA silencing by a geminivirus nuclear protein, AC2, correlates with transactivation of host genes. J Virol 2005, 79: 2517-2527.Senthil G, Liu H, Puram VG, Clark A, Stromberg A, Goodin MM: Specific and common changes in Nicotiana benthamiana gene expression in response to infection by enveloped viruses. J Gen Virol 2005, 86: 2615-2625.Marathe R, Guan Z, Anandalakshmi R, Zhao H, Dinesh-Kumar SP: Study of Arabidopsis thaliana resistome in response to cucumber mosaic virus infection using whole genome microarray. Plant Mol Biol 2004, 55: 501-520.Agudelo-Romero P, Carbonell P, de la Iglesia F, Carrera J, Rodrigo G, Jaramillo A, Perez-Amador MA, Elena SF: Changes in the gene expression profile of Arabidopsis thaliana after infection with Tobacco etch virus. Virol J 2008, 5: 92.Itaya A, Matsuda Y, Gonzales RA, Nelson RS, Ding B: Potato spindle tuber viroid strains of different pathogenicity induces and suppresses expression of common and unique genes in infected tomato. Mol Plant Microbe Interact 2002, 15: 990-999.Huang Z, Yeakley JM, Garcia EW, Holdridge JD, Fan JB, Whitham SA: Salicylic acid-dependent expression of host genes in compatible Arabidopsis-virus interactions. Plant Physiol 2005, 137: 1147-1159.Rizza S, Conesa A, Juarez J, Catara A, Navarro L, Duran-Vila N, Ancillo G: Microarray analysis of Etrog citron (Citrus medica L.) reveals changes in chloroplast, cell wall, peroxidase and symporter activities in response to viroid infection. Mol Plant Pathol 2012,13(8):852-864.Golem S, Culver JN: Tobacco mosaic virus induced alterations in the gene expression profile of Arabidopsis thaliana. Mol Plant Microbe Interact 2003, 16: 681-688.Dardick C: Comparative expression profiling of Nicotiana benthamiana leaves systemically infected with three fruit tree viruses. Mol Plant Microbe Interact 2007, 20: 1004-1017.Hull R: In Matthews’ Plant Virology. London: Edited by Academic Press; 2002.Gonzalez-Jara P, Tenllado F, Martinez-Garcia B, Atencio FA, Barajas D, Vargas M, Diaz-Ruiz J, Diaz-Ruiz JR: Host-dependent differences during synergistic infection by Potyviruses with potato virus X. Mol Plant Pathol 2004, 5: 29-35.Gonzalez-Jara P, Atencio FA, Martinez-Garcia B, Barajas D, Tenllado F, Diaz-Ruiz JR: A Single Amino Acid Mutation in the Plum pox virus Helper Component-Proteinase Gene Abolishes Both Synergistic and RNA Silencing Suppression Activities. Phytopathology 2005, 95: 894-901.Vance VB: Replication of potato virus X RNA is altered in coinfections with potato virus Y. Virology 1991, 182: 486-494.Garcia-Marcos A, Pacheco R, Martianez J, Gonzalez-Jara P, Diaz-Ruiz JR, Tenllado F: Transcriptional changes and oxidative stress associated with the synergistic interaction between Potato virus X and Potato virus Y and their relationship with symptom expression. Mol Plant Microbe Interact 2009, 22: 1431-1444.Postnikova OA, Nemchinov LG: Comparative analysis of microarray data in Arabidopsis transcriptome during compatible interactions with plant viruses. Virol J 2012, 9: 101.Zanchin A, Bonghi C, Casadoro G, Ramina A, Rascio N: Cell enlargement and cell separation during peach fruit development. International Journal of Plant Science 1994, 155: 49-56.Herranz MC, Sanchez-Navarro JA, Aparicio F, Pallas V: Simultaneous detection of six stone fruit viruses by non-isotopic molecular hybridization using a unique riboprobe or ‘polyprobe’. J Virol Methods 2005, 124: 49-55.Pallas V, Mas P, Sanchez-Navarro JA: Detection of plant RNA viruses by nonisotopic dot-blot hybridization. Methods Mol Biol 1998, 81: 461-468.Lilly ST, Drummond RS, Pearson MN, MacDiarmid RM: Identification and validation of reference genes for normalization of transcripts from virus-infected Arabidopsis thaliana. Mol Plant Microbe Interact 2011, 24: 294-304.Cosgrove JD: Expansive growth of plant cell walls. Plant Physiol Biochem 2000,38(1–2):109-124.Tessitori M, Maria G, Capasso C, Catara G, Rizza S, De Luca V, Catara A, Capasso A, Carginale V: Differential display analysis of gene expression in Etrog citron leaves infected by Citrus viroid III. Biochim Biophys Acta 2007, 1769: 228-235.Rizza S, Capasso C, Catara A, Capasso A, Conte E, Catara A Proceedings of the 17th Conference of the International Organization of Citrus Virologists-IOCV, pp. XVII. In Transcriptional response of Troyer citrange, sour orange and alemow rootstocks to Citrus viroid IIIb (CVd-IIIb) infection. Adana, Turkey: Conference of the International Organization of Citrus Virologists; 2010:142-149. http://www.ivia.es/iocv/Owens RA, Tech KB, Shao JY, Sano T, Baker CJ: Global analysis of tomato gene expression during Potato spindle tuber viroid infection reveals a complex array of changes affecting hormone signaling. Mol Plant Microbe Interact 2012, 25: 582-598.Ogundiwin EA, Marti C, Forment J, Pons C, Granell A, Gradziel TM, Peace CP, Crisosto CH: Development of ChillPeach genomic tools and identification of cold-responsive genes in peach fruit. Plant Mol Biol 2008, 68: 379-397.Sánchez-Navarro JA FA, Rowhani A, Pallás V: Comparative analysis of ELISA, nonradioactive molecular hybridization and PCR for the detection of Prunus necrotic ringspot virus in herbaceous and prunus host. Plant Pathol 1998, 47: 780-786.Astruc N, Marcos JF, Macquaire G, Candresse T, Pallas V: Studies on the diagnosis of hop stunt viroid in fruit trees: Identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents. Eur J Plant Pathol 1996, 102: 837-846.Myrta A, Di Terlizzi B, Pallas V, Savino V: Viruses and viroids of apricot in the Mediterranean: incidence and biodiversity. Acta Horticulturae 2006, 701: 409-417.Bouzayen M, Latché A, Nath P, Pech JC: Mechanism of fruit ripening. In Plant Developmental Biology- Biotechnological Perspectives: Volume I Edited by: Pua EC, Darvey MR. 2010, 319-339. Chapter 16Trainotti L, Bonghi C, Ziliotto F, Zanin D, Rasori A, Casadoro G, Ramina A, T P: The use of microarray mPEACH 1.0 to investigate transcriptome changes during transition from pre-climateric to climacteric phase in peach fruit. Plant Sci 2006, 170: 606-613.Lombardo VA, Osorio S, Borsani J, Lauxmann MA, Bustamante CA, Budde CO, Andreo CS, Lara MV, Fernie AR MFD: Metabolic profiling during peach fruit development and ripening reveals the metabolic networks that underpin each developmental stage. Plant Physiol 2011,157(4):1696-1710.Manganaris GA RA, Bassi D, Geuna F, Ramina A, Tonutti P, Bonghi C: Comparative transcript profiling of apricot (Prunus armeniaca L.) fruit development and on-tree ripening. Tree Genet Genomes 2011, 7: 609-616.Uyemoto JK, Scott SW: Important diseases of Prunus caused by viruses and other graft-transmissible pathogens in California and South Carolina. Plant Dis 1992, 76: 5-11.Li J, Yang H, Peer WA, Richter G, Blakeslee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S, Khodakovskaya M, Richards EL, et al.: Arabidopsis H+-PPase AVP1 regulates auxin-mediated organ development. Science 2005, 310: 121-125.Paponov IA, Paponov M, Teale W, Menges M, Chakrabortee S, Murray JA, Palme K: Comprehensive transcriptome analysis of auxin responses in Arabidopsis. Mol Plant 2008, 1: 321-337.Padmanabhan MS, Goregaoker SP, Golem S, Shiferaw H, Culver JN: Interaction of the tobacco mosaic virus replicase protein with the Aux/IAA protein PAP1/IAA26 is associated with disease development. J Virol 2005, 79: 2549-2558.Padmanabhan MS, Shiferaw H, Culver JN: The Tobacco mosaic virus replicase protein disrupts the localization and function of interacting Aux/IAA proteins. Mol Plant Microbe Interact 2006, 19: 864-873.Padmanabhan MS, Kramer SR, Wang X, Culver JN: Tobacco mosaic virus replicase-auxin/indole acetic acid protein interactions: reprogramming the auxin response pathway to enhance virus infection. J Virol 2008, 82: 2477-2485.Kuhn JM, Boisson-Dernier A, Dizon MB, Maktabi MH, Schroeder JI: The protein phosphatase AtPP2CA negatively regulates abscisic acid signal transduction in Arabidopsis, and effects of abh1 on AtPP2CA mRNA. Plant Physiol 2006, 140: 127-139.Whenham RJ, Fraser RSS, Brown LP, Payne JA: Tobacco-mosaic-virus-induced increase in abscisic-acid concentration in tobacco leaves: Intracellular location in light and dark-green areas, and relationship to symptom development. Planta 1986, 168: 592-598.Bari R, Jones JD: Role of plant hormones in plant defence responses. Plant Mol Biol 2009, 69: 473-488.Kotchoni SO, Kuhns C, Ditzer A, Kirch HH, Bartels D: Over-expression of different aldehyde dehydrogenase genes in Arabidopsis thaliana confers tolerance to abiotic stress and protects plants against lipid peroxidation and oxidative stress. Plant Cell Environ 2006, 29: 1033-1048.Mowla SB, Cuypers A, Driscoll SP, Kiddle G, Thomson J, Foyer CH, Theodoulou FL: Yeast complementation reveals a role for an Arabidopsis thaliana late embryogenesis abundant (LEA)-like protein in oxidative stress tolerance. Plant J 2006, 48: 743-756.Amari K, Diaz-Vivancos P, Pallas V, Sanchez-Pina MA, Hernandez JA: Oxidative stress induction by Prunus necrotic ringspot virus infection in apricot seeds. Physiol Plant 2007, 131: 302-310.Gilroy EM, Hein I, van der Hoorn R, Boevink PC, Venter E, McLellan H, Kaffarnik F, Hrubikova K, Shaw J, Holeva M, et al.: Involvement of cathepsin B in the plant disease resistance hypersensitive response. Plant J 2007, 52: 1-13.Kruger J, Thomas CM, Golstein C, Dixon MS, Smoker M, Tang S, Mulder L, Jones JD: A tomato cysteine protease required for Cf-2-dependent disease resistance and suppression of autonecrosis. Science 2002, 296: 744-747.Bernoux M, Timmers T, Jauneau A, Briere C, De Wit PJ, Marco Y, Deslandes L: RD19, an Arabidopsis cysteine protease required for RRS1-R-mediated resistance, is relocalized to the nucleus by the Ralstonia solanacearum PopP2 effector. Plant Cell 2008, 20: 2252-2264.Shabab M, Shindo T, Gu C, Kaschani F, Pansuriya T, Chintha R, Harzen A, Colby T, Kamoun S, van der Hoorn RA: Fungal effector protein AVR2 targets diversifying defense-related cys proteases of tomato. Plant Cell 2008, 20: 1169-1183.Van Esse HP, Van’t Klooster JW, Bolton MD, Yadeta KA, Van Baarlen P, Boeren S, Vervoort J, De Wit PJ, Thomma BP: The Cladosporium fulvum virulence protein Avr2 inhibits host proteases required for basal defense. Plant Cell 2008, 20: 1948-1963.Song J, Win J, Tian M, Schornack S, Kaschani F, Ilyas M, van der Hoorn RA, Kamoun S: Apoplastic effectors secreted by two unrelated eukaryotic plant pathogens target the tomato defense protease Rcr3. Proc Natl Acad Sci U S A 2009, 106: 1654-1659.Tian M, Win J, Song J, van der Hoorn R, van der Knaap E, Kamoun S: A Phytophthora infestans cystatin-like protein targets a novel tomato papain-like apoplastic protease. Plant Physiol 2007, 143: 364-377.Rooney H, Van’t Klooster J, Van der Hoorn R, Joosten M, Jones J: Cladosporium Avr2 inhibits tomato Rcr3 protease required for Cf-2-dependent disease resistance. Science 2005, 308: 1783-1786.Auger AJ: Tomato ringspot virus associated with brownline disease on prune trees in Chile. Acta Horticulturae 1989, 235: 197-204.Herrera G: Enfermedades causadas por virus en frutales en Chile. Santiago, Chile: Instituto de Investigación Agropecuaria; 2001. Boletín INIA N°52. 65pFiore N, Abou Ghanem-Sabanadzovic N, Infante R, Myrta A, Pallás V: Detection of Peach latent mosaic viroid in stone fruits from Chile. In Option Méditerranéennes, Sér. B/n°45 –Virus ad virus-like disease of stone fruits, with particular reference to the Mediterranean region Edited by: Myrta A, Di Terlizzi B, Savino V. 2003, 143-145.Torres H, Gómez G, Pallás V, Stamo B, Shalaby A, Aouane B, Gavriel I, Kominek P, Caglayan K, Sipahioglu M, et al.: Detection by tissue printing of stone fruit viroids, from europe, the mediterranean and north and south America. Acta Horticulturae 2004, 657: 379-383.Peiró A, Pallás V, Sánchez-Navarro JA: Simultaneous detection of eight viruses and two viroids affecting stone fruit trees by using a unique polyprobe. Eur J Plant Pathol 2012,132(4):469-475.Meisel L, Fonseca B, Gonzalez S, Baeza-Yates R, Cambiazo V, Campos R, Gonzalez M, Orellana A, Retamales J, Silva H: A rapid and efficient method for purifying high quality total RNA from peaches (Prunus persica) for functional genomics analyses. Biol Res 2005, 38: 83-88.Van Gelder RN, Von Zastrow ME, Yool A, Dement WC, Barchas JD JHE: Amplified RNA synthesized from limited quantities of heterogeneous cDNA. Proc Natl Acad Sci U S A 1990,87(5):1663-1667.Tusher VG, Tibshirani R, Chu G: Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 2001, 98: 5116-5121.Sanchez-Navarro JA, Canizares MC, Cano EA, Pallas V: Simultaneous detection of five carnation viruses by non-isotopic molecular hybridization. J Virol Methods 1999, 82: 167-175

Comparative analysis among the small RNA populations of source, sink and conductive tissues in two different plant-virus pathosystems

Conclusions: We compare for the first time the sRNA profile of four different tissues, including source, sink and conductive (phloem) tissues, in two plant-virus pathosystems. Our results indicate that antiviral silencing machinery in melon and cucumber acts mainly through DCL4. Upon infection, the total sRNA pattern in phloem remains unchanged in contrast to the rest of the analyzed tissues indicating a certain tissue-tropism to this polulation. Independently of the accumulation level of the vsRNAs both viruses were able to modulate the host sRNA pattern.We thank Dr A. Niehl for critical reading and helpful comments on the manuscript. This work was funded by a supporting program for the research from the Universidad Politecnica de Valencia (PAID-05-10), a grant BIO2011-25018 from the Spanish granting agency Direccion General de Investigacion Cientifica and the PROMETEO program 2011/003 from the Generalitat Valenciana. MCH is the recipient of a contract from JAE-DOC program of the CSIC, JAN is the recipient of a postdoctoral contract from the Ministerio de Educacion y Ciencia of Spain.Herranz Gordo, MDC.; Navarro Bohigues, JA.; Sommen, E.; Pallás Benet, V. (2015). Comparative analysis among the small RNA populations of source, sink and conductive tissues in two different plant-virus pathosystems. BMC Genomics. 16:1-15. https://doi.org/10.1186/s12864-015-1327-5S11516Pumplin N, Voinnet O. RNA silencing suppression by plant pathogens: defence, counter-defence and counter-counter-defence. Nat Rev Microbiol. 2013;11(11):745–60.Brodersen P, Voinnet O. The diversity of RNA silencing pathways in plants. Trends Genet. 2006;22(5):268–80.Ghildiyal M, Zamore PD. Small silencing RNAs: an expanding universe. Nat Rev Genet. 2009;10(2):94–108.Ciaudo C, Jay F, Okamoto I, Chen CJ, Sarazin A, Servant N, et al. RNAi-dependent and independent control of LINE1 accumulation and mobility in mouse embryonic stem cells. PLoS Genet. 2013;9(11):e1003791.Ding SW, Voinnet O. Antiviral immunity directed by small RNAs. Cell. 2007;130(3):413–26.Szittya G, Moxon S, Pantaleo V, Toth G, Rusholme Pilcher RL, Moulton V, et al. Structural and functional analysis of viral siRNAs. PLoS Pathog. 2010;6(4):e1000838.Donaire L, Wang Y, Gonzalez-Ibeas D, Mayer KF, Aranda MA, Llave C. Deep-sequencing of plant viral small RNAs reveals effective and widespread targeting of viral genomes. Virology. 2009;392(2):203–14.Voinnet O. Origin, biogenesis, and activity of plant microRNAs. Cell. 2009;136(4):669–87.Liu Q, Feng Y, Zhu Z. Dicer-like (DCL) proteins in plants. Funct Integr Genomics. 2009;9(3):277–86.Henderson IR, Zhang X, Lu C, Johnson L, Meyers BC, Green PJ, et al. Dissecting Arabidopsis thaliana DICER function in small RNA processing, gene silencing and DNA methylation patterning. Nat Genet. 2006;38(6):721–5.Margis R, Fusaro AF, Smith NA, Curtin SJ, Watson JM, Finnegan EJ, et al. The evolution and diversification of Dicers in plants. FEBS Lett. 2006;580(10):2442–50.Deleris A, Gallego-Bartolome J, Bao J, Kasschau KD, Carrington JC, Voinnet O. Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense. Science. 2006;313(5783):68–71.Blevins T, Rajeswaran R, Shivaprasad PV, Beknazariants D, Si-Ammour A, Park HS, et al. Four plant Dicers mediate viral small RNA biogenesis and DNA virus induced silencing. Nucleic Acids Res. 2006;34(21):6233–46.Bouche N, Lauressergues D, Gasciolli V, Vaucheret H. An antagonistic function for Arabidopsis DCL2 in development and a new function for DCL4 in generating viral siRNAs. EMBO J. 2006;25(14):3347–56.Moissiard G, Voinnet O. RNA silencing of host transcripts by cauliflower mosaic virus requires coordinated action of the four Arabidopsis Dicer-like proteins. Proc Natl Acad Sci U S A. 2006;103(51):19593–8.Qu F, Ye X, Morris TJ. Arabidopsis DRB4, AGO1, AGO7, and RDR6 participate in a DCL4-initiated antiviral RNA silencing pathway negatively regulated by DCL1. Proc Natl Acad Sci U S A. 2008;105(38):14732–7.Vaucheret H. Plant ARGONAUTES. Trends Plant Sci. 2008;13(7):350–8.Hutvagner G, Simard MJ. Argonaute proteins: key players in RNA silencing. Nat Rev Mol Cell Biol. 2008;9(1):22–32.Voinnet O. Use, tolerance and avoidance of amplified RNA silencing by plants. Trends Plant Sci. 2008;13(7):317–28.Palauqui JC, Elmayan T, Pollien JM, Vaucheret H. Systemic acquired silencing: transgene-specific post-transcriptional silencing is transmitted by grafting from silenced stocks to non-silenced scions. EMBO J. 1997;16(15):4738–45.Yoo BC, Kragler F, Varkonyi-Gasic E, Haywood V, Archer-Evans S, Lee YM, et al. A systemic small RNA signaling system in plants. Plant Cell. 2004;16(8):1979–2000.Buhtz A, Pieritz J, Springer F, Kehr J. Phloem small RNAs, nutrient stress responses, and systemic mobility. BMC Plant Biol. 2010;10:64.Buhtz A, Springer F, Chappell L, Baulcombe DC, Kehr J. Identification and characterization of small RNAs from the phloem of Brassica napus. Plant J. 2008;53(5):739–49.Rodriguez-Medina C, Atkins CA, Mann AJ, Jordan ME, Smith PM. Macromolecular composition of phloem exudate from white lupin (Lupinus albus L.). BMC Plant Biol. 2011;11:36.Pallas V, Gomez G. Phloem RNA-binding proteins as potential components of the long-distance RNA transport system. Frontiers in Plant Science. 2013;4:130.Tournier B, Tabler M, Kalantidis K. Phloem flow strongly influences the systemic spread of silencing in GFP Nicotiana benthamiana plants. Plant J. 2006;47(3):383–94.Hamilton A, Voinnet O, Chappell L, Baulcombe D. Two classes of short interfering RNA in RNA silencing. EMBO J. 2002;21(17):4671–9.Voinnet O. MicroRNA and autophagy--C. elegans joins the crew. EMBO Rep. 2013;14(6):485–7.Dunoyer P, Schott G, Himber C, Meyer D, Takeda A, Carrington JC, et al. Small RNA duplexes function as mobile silencing signals between plant cells. Science. 2010;328(5980):912–6.Brosnan CA, Mitter N, Christie M, Smith NA, Waterhouse PM, Carroll BJ. Nuclear gene silencing directs reception of long-distance mRNA silencing in Arabidopsis. Proc Natl Acad Sci U S A. 2007;104(37):14741–6.Silva TF, Romanel EA, Andrade RR, Farinelli L, Osteras M, Deluen C, et al. Profile of small interfering RNAs from cotton plants infected with the polerovirus Cotton leafroll dwarf virus. BMC Mol Biol. 2011;12:40.Martinez G, Donaire L, Llave C, Pallas V, Gomez G. High-throughput sequencing of Hop stunt viroid-derived small RNAs from cucumber leaves and phloem. Mol Plant Pathol. 2010;11(3):347–59.Donaire L, Barajas D, Martinez-Garcia B, Martinez-Priego L, Pagan I, Llave C. Structural and genetic requirements for the biogenesis of tobacco rattle virus-derived small interfering RNAs. J Virol. 2008;82(11):5167–77.Qi X, Bao FS, Xie Z. Small RNA deep sequencing reveals role for Arabidopsis thaliana RNA-dependent RNA polymerases in viral siRNA biogenesis. PLoS One. 2009;4(3):e4971.Pantaleo V, Saldarelli P, Miozzi L, Giampetruzzi A, Gisel A, Moxon S, et al. Deep sequencing analysis of viral short RNAs from an infected Pinot Noir grapevine. Virology. 2010;408(1):49–56.Lin KY, Cheng CP, Chang BC, Wang WC, Huang YW, Lee YS, et al. Global analyses of small interfering RNAs derived from Bamboo mosaic virus and its associated satellite RNAs in different plants. PLoS One. 2010;5(8):e11928.Navarro B, Pantaleo V, Gisel A, Moxon S, Dalmay T, Bisztray G, et al. Deep sequencing of viroid-derived small RNAs from grapevine provides new insights on the role of RNA silencing in plant-viroid interaction. PLoS One. 2009;4(11):e7686.Martin R, Arenas C, Daros JA, Covarrubias A, Reyes JL, Chua NH. Characterization of small RNAs derived from Citrus exocortis viroid (CEVd) in infected tomato plants. Virology. 2007;367(1):135–46.St-Pierre P, Hassen IF, Thompson D, Perreault JP. Characterization of the siRNAs associated with peach latent mosaic viroid infection. Virology. 2009;383(2):178–82.Di Serio F, Gisel A, Navarro B, Delgado S, de Alba AE M, Donvito G, et al. Deep sequencing of the small RNAs derived from two symptomatic variants of a chloroplastic viroid: implications for their genesis and for pathogenesis. PLoS One. 2009;4(10):e7539.Li R, Gao S, Hernandez AG, Wechter WP, Fei Z, Ling KS. Deep sequencing of small RNAs in tomato for virus and viroid identification and strain differentiation. PLoS One. 2012;7(5):e37127.Hu Q, Hollunder J, Niehl A, Korner CJ, Gereige D, Windels D, et al. Specific impact of tobamovirus infection on the Arabidopsis small RNA profile. PLoS One. 2011;6(5):e19549.Hibi T, Furuki I. Melon Necrotic Spot Virus. In: CMI: AAB Descriptions of Plants Viruses N° 302. Kew, UK: Commonwealth Mycological Institute; 1985.Riviere CJ, Rochon DM. Nucleotide sequence and genomic organization of melon necrotic spot virus. J Gen Virol. 1990;71(Pt 9):1887–96.Diaz JA, Nieto C, Moriones E, Truniger V, Aranda MA. Molecular characterization of a Melon necrotic spot virus strain that overcomes the resistance in melon and nonhost plants. Mol Plant Microbe Interact. 2004;17(6):668–75.Navarro JA, Genoves A, Climent J, Sauri A, Martinez-Gil L, Mingarro I, et al. RNA-binding properties and membrane insertion of Melon necrotic spot virus (MNSV) double gene block movement proteins. Virology. 2006;356(1–2):57–67.Genoves A, Navarro JA, Pallas V. A self-interacting carmovirus movement protein plays a role in binding of viral RNA during the cell-to-cell movement and shows an actin cytoskeleton dependent location in cell periphery. Virology. 2009;395(1):133–42.Genoves A, Navarro JA, Pallas V. The Intra- and intercellular movement of Melon necrotic spot virus (MNSV) depends on an active secretory pathway. Mol Plant Microbe Interact. 2010;23(3):263–72.Serra-Soriano M, Pallas V, Navarro JA. A model for transport of a viral membrane protein through the early secretory pathway: minimal sequence and endoplasmic reticulum lateral mobility requirements. Plant J. 2014;77(6):863–79.Genoves A, Navarro JA, Pallas V. Functional analysis of the five melon necrotic spot virus genome-encoded proteins. J Gen Virol. 2006;87(Pt 8):2371–80.Pallas V, Aparicio F, Herranz MC, Amari K, Sanchez-Pina MA, Myrta A, et al. Ilarviruses of Prunus spp.: a continued concern for fruit trees. Phytopathology. 2012;102(12):1108–20.Pallas V, Aparicio F, Herranz MC, Sanchez-Navarro JA, Scott SW. The molecular biology of ilarviruses. Adv Virus Res. 2013;87:139–81.Varkonyi-Gasic E, Wu R, Wood M, Walton EF, Hellens RP. Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant Methods. 2007;3:12.Blevins T, Rajeswaran R, Aregger M, Borah BK, Schepetilnikov M, Baerlocher L, et al. Massive production of small RNAs from a non-coding region of Cauliflower mosaic virus in plant defense and viral counter-defense. Nucleic Acids Res. 2011;39(12):5003–14.Takeda A, Tsukuda M, Mizumoto H, Okamoto K, Kaido M, Mise K, et al. A plant RNA virus suppresses RNA silencing through viral RNA replication. EMBO J. 2005;24(17):3147–57.Andersson MG, Haasnoot PC, Xu N, Berenjian S, Berkhout B, Akusjarvi G. Suppression of RNA interference by adenovirus virus-associated RNA. J Virol. 2005;79(15):9556–65.Himeno M, Maejima K, Komatsu K, Ozeki J, Hashimoto M, Kagiwada S, et al. Significantly low level of small RNA accumulation derived from an encapsidated mycovirus with dsRNA genome. Virology. 2010;396(1):69–75.Aparicio F, Vilar M, Perez-Paya E, Pallas V. The coat protein of prunus necrotic ringspot virus specifically binds to and regulates the conformation of its genomic RNA. Virology. 2003;313(1):213–23.Ruiz-Ruiz S, Navarro B, Gisel A, Pena L, Navarro L, Moreno P, et al. Citrus tristeza virus infection induces the accumulation of viral small RNAs (21-24-nt) mapping preferentially at the 3′-terminal region of the genomic RNA and affects the host small RNA profile. Plant Mol Biol. 2011;75(6):607–19.Folimonova SY, Folimonov AS, Tatineni S, Dawson WO. Citrus tristeza virus: survival at the edge of the movement continuum. J Virol. 2008;82(13):6546–56.Kreuze JF, Perez A, Untiveros M, Quispe D, Fuentes S, Barker I, et al. Complete viral genome sequence and discovery of novel viruses by deep sequencing of small RNAs: a generic method for diagnosis, discovery and sequencing of viruses. Virology. 2009;388(1):1–7.Karyeija RF, Kreuze JF, Gibson RW, Valkonen JP. Synergistic interactions of a potyvirus and a phloem-limited crinivirus in sweet potato plants. Virology. 2000;269(1):26–36.Melnyk CW, Molnar A, Bassett A, Baulcombe DC. Mobile 24 nt small RNAs direct transcriptional gene silencing in the root meristems of Arabidopsis thaliana. Curr Biol. 2011;21(19):1678–83.Gosalvez-Bernal B, Genoves A, Navarro JA, Pallas V, Sanchez-Pina MA. Distribution and pathway for phloem-dependent movement of Melon necrotic spot virus in melon plants. Mol Plant Pathol. 2008;9(4):447–61.Harper SJ, Cowell SJ, Robertson CJ, Dawson WO. Differential tropism in roots and shoots infected by Citrus tristeza virus. Virology. 2014;460–461:91–9.Andika IB, Kondo H, Tamada T. Evidence that RNA silencing-mediated resistance to beet necrotic yellow vein virus is less effective in roots than in leaves. Mol Plant Microbe Interact. 2005;18(3):194–204.Mi S, Cai T, Hu Y, Chen Y, Hodges E, Ni F, et al. Sorting of small RNAs into Arabidopsis argonaute complexes is directed by the 5' terminal nucleotide. Cell. 2008;133(1):116–27.Takeda A, Iwasaki S, Watanabe T, Utsumi M, Watanabe Y. The mechanism selecting the guide strand from small RNA duplexes is different among argonaute proteins. Plant Cell Physiol. 2008;49(4):493–500.Wu L, Zhang Q, Zhou H, Ni F, Wu X, Qi Y. Rice MicroRNA effector complexes and targets. Plant Cell. 2009;21(11):3421–35.Xu Y, Huang L, Fu S, Wu J, Zhou X. Population diversity of rice stripe virus-derived siRNAs in three different hosts and RNAi-based antiviral immunity in Laodelphgax striatellus. PLoS One. 2012;7(9):e46238

Adelante / Endavant

Séptimo desafío por la erradicación de la violencia contra las mujeres del Institut Universitari d’Estudis Feministes i de Gènere "Purificación Escribano" de la Universitat Jaume

Suppression of the yeast <i>sec12-1</i> by the plant AtSec12 homologue.

<p>A) Total protein extracts of yeast wild-type (WT) and <i>sec12-1</i> mutant cells untransformed or transformed with the empty vector (pVV) or expressing plant <i>AtSEC12</i> were resolved by SDS-PAGE and immunoblotted with anti-AtSec12 antibodies. B) Yeast cell cultures grown to mid-exponential phase at 25°C were spotted on YPD medium containing <i>MATa bar1</i> mutant cells to determine α-factor secretion at permissive (25°C) and various restrictive temperatures (30°C, 35°C and 37°C). C) Localization of a COPII cargo the SNARE GFP-Snc1 in the <i>sec12-1</i> mutant transformed with either pVV (empty) or AtSec12 plasmids was determined by epifluorescence observation (with GFP and Differential interference contrast (DIC) filters) on cell cultures grown to mid-exponential phase at 25°C. D) Total protein extracts from wild-type (WT) cells or <i>sec12-1</i> mutant cells transformed with pVV or AtSEC12 plasmid and with GFP-Snc1 vector and grown at 25°C were resolved by SDS-PAGE and immunoblotted with anti-GFP antibodies.</p

The <i>Arabidopsis thaliana</i> AtSar1D complements the secretory defect of the yeast <i>sar1-2</i> mutant cells.

<p>A) Total protein extracts of WT and <i>sar1-2</i> mutant cells untransformed or transformed with empty vector (pVV) or vectors bearing the plant <i>AtSAR1</i> isoforms A to D grown at 25°C were resolved by SDS-PAGE and immunoblotted with anti-AtSar1B antibodies. B) The same cell cultures as in A) were spotted on YPD medium containing <i>MATa bar1</i> mutant cells to analyze α-factor secretion at permissive (25°C) and various restrictive temperatures (30°C and 35°C). C) The <i>sar1-2</i> mutant cells transformed with the empty (pVV) or the AtSar1D plasmid were co-transformed with the GFP-Snc1 vector; these cells were grown at 25°C to mid-exponential phase prior their observation by fluorescence microscopy. D) Total proteins were extracted from the same cell cultures as in C) and resolved by SDS-PAGE prior immunoblotting with anti-GFP antibodies to detect the GFP-Snc1 proteins.</p

Complementation of the yeast <i>sec13-1</i> mutant by the <i>Arabidopsis</i> thaliana AtSec13A isoform.

<p>A) Yeast wild-type (WT) and <i>sec13-1</i> mutant cells untransformed or transformed with empty vector (pVV) or bearing plant AtSec13A or AtSec13B isoforms were grown at 25°C to mid-exponential phase prior lysis to extract total proteins that were subjected to western-blot analysis with anti-AtSec13A antibodies. B) The same cell cultures at in A) were spotted on YPD medium containing <i>MATa bar1</i> mutant cells to determine α-factor secretion at permissive (25°C) and different restrictive temperatures (30°C, 35°C and 37°C). C) Localization of the COPII cargo GFP-Snc1 in the <i>sec13-1</i> mutant transformed with either the empty (pVV) or AtSec13A plasmid was determined by epifluorescence on mid-exponential phase cell cultures at permissive (25°C) or after a 2 h shift at restrictive (37°C) temperature. D) Total proteins extracted from the same strains as in C) but after a 4 h shift at 37°C were resolved by SDS-PAGE and immunoblotted with anti-GFP antibodies to determine the phosphorylation status of the GFP-Snc1 proteins.</p