The RNA-binding protein, ZFP36L2, influences ovulation and oocyte maturation

ZFP36L2 protein destabilizes AU-rich element-containing transcripts and has been implicated in female fertility. In the C57BL/6NTac mouse, a mutation in Zfp36l2 that results in the decreased expression of a form of ZFP36L2 in which the 29 N-terminal amino acid residues have been deleted, ΔN-ZFP36L2, leads to fertilized eggs that arrest at the two-cell stage. Interestingly, homozygous ΔN-Zfp36l2 females in the C57BL/6NTac strain release 40% fewer eggs than the WT littermates (Ramos et al., 2004), suggesting an additional defect in ovulation and/or oocyte maturation. Curiously, the same ΔN-Zfp36l2 mutation into the SV129 strain resulted in anovulation, prompting us to investigate a potential problem in ovulation and oocyte maturation. Remarkably, only 20% of ΔN-Zfp36l2 oocytes in the 129S6/SvEvTac strain matured ex vivo, suggesting a defect on the oocyte meiotic maturation process. Treatment of ΔN-Zfp36l2 oocytes with a PKA inhibitor partially rescued the meiotic arrested oocytes. Furthermore, cAMP levels were increased in ΔN-Zfp36l2 oocytes, linking the cAMP/PKA pathway and ΔN-Zfp36l2 with meiotic arrest. Since ovulation and oocyte maturation are both triggered by LHR signaling, the downstream pathway was investigated. Adenylyl cyclase activity was increased in ΔN-Zfp36l2 ovaries only upon LH stimulation. Moreover, we discovered that ZFP36L2 interacts with the 3′UTR of LHR mRNA and that decreased expression levels of Zfp36l2 correlates with higher levels of LHR mRNA in synchronized ovaries. Furthermore, overexpression of ZFP36L2 decreases the endogenous expression of LHR mRNA in a cell line. Therefore, we propose that lack of the physiological down regulation of LHR mRNA levels by ZFP36L2 in the ovaries is associated with anovulation and oocyte meiotic arrest.Fil: Ball, Christopher B.. University of North Carolina; Estados UnidosFil: Rodriguez, Karina F.. National Institutes of Health; Estados UnidosFil: Stumpo, Deborah J.. National Institutes of Health; Estados UnidosFil: Ribeiro Neto, Fernando. National Institutes of Health; Estados UnidosFil: Korach, Kenneth S.. National Institutes of Health; Estados UnidosFil: Blackshear, Perry J.. University of Duke; Estados Unidos. National Institutes of Health; Estados UnidosFil: Birnbaumer, Lutz. National Institutes of Health; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ramos, Silvia B. V.. University of North Carolina; Estados Unido

Bulk and single-molecule analysis of a bacterial DNA2-like helicase-nuclease reveals a single-stranded DNA looping motor

DNA2 is an essential enzyme involved in DNA replication and repair in eukaryotes. In a search for homologues of this protein, we identified and characterised Geobacillus stearothermophilus Bad, a bacterial DNA helicase-nuclease with similarity to human DNA2. We show that Bad contains an Fe-S cluster and identify four cysteine residues that are likely to co-ordinate the cluster by analogy to DNA2. The purified enzyme specifically recognises ss-dsDNA junctions and possesses ssDNA-dependent ATPase, ssDNA binding, ssDNA endonuclease, 5' to 3' ssDNA translocase and 5' to 3' helicase activity. Single molecule analysis reveals that Bad is a processive DNA motor capable of moving along DNA for distances of >4 kb at a rate of ∼200 bp per second at room temperature. Interestingly, as reported for the homologous human and yeast DNA2 proteins, the DNA unwinding activity of Bad is cryptic and can be unmasked by inactivating the intrinsic nuclease activity. Strikingly, our experiments show that the enzyme loops DNA while translocating, which is an emerging feature of processive DNA unwinding enzymes. The bacterial Bad enzymes will provide an excellent model system for understanding the biochemical properties of DNA2-like helicase-nucleases and DNA looping motor proteins in general.Wellcome Trust [100401/Z/12/Z to M.D.]; EuropeanResearch Council [681299 to F.M.-H.]; Spanish Min-istry of Economy and Competitiveness [BFU2017-83794-PAEI/FEDER, UE to F.M.-H.]; Comunidad de MadridTec4Bio [S2018/NMT-4443 to F.M.-H.]; NanoBioCancer[Y2018/BIO-4747 to F.M.-H.]. Funding for open accesscharge: Wellcome Trust [100401/Z/12/Z].Peer reviewe

Gadget for anchovy 9a South: Model description and results to provide catch advice and reference points (WGHANSA-1 2021)

The model speci fications presented below correspond to those benchmarked in WKPELA 2018. The main difference is that results are presented now for the end of the second quarter of each year instead of being presented at the end of the fourth quarter. This responds to practical modi cations in the de nition of the assessment year, now it goes from July 1st to June 30th of the next year. Model speci fications for this year are presented in section 2.2 and ??, as well as estimated parameters after optimization in Table 2

Gadget for anchovy 9a South: Model description and results to provide catch advice and reference points (WGHANSA-1 2022).

The model speci fications presented below correspond to those benchmarked in WKPELA 2018. The main difference is that results are presented now for the end of the second quarter of each year instead of be presented at the end of the fourth quarter. This responds to practical modi cations in the defi nition of the assessment year, now it goes from July 1st to June 30th of the next year. Specifi c model assumptions for this year are presented in section 2.2 and 3, as well as estimated parameters after optimization in Table 2

Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA

Human DNA helicase B (HELB) is a poorly characterized helicase suggested to play both positive and negative regulatory roles in DNA replication and recombination. In this work, we used bulk and single-molecule approaches to characterize the biochemical activities of HELB protein with a particular focus on its interactions with Replication Protein A (RPA) and RPA–single-stranded DNA (ssDNA) filaments. HELB is a monomeric protein that binds tightly to ssDNA with a site size of ∼20 nucleotides. It couples ATP hydrolysis to translocation along ssDNA in the 5′ to 3′ direction accompanied by the formation of DNA loops. HELB also displays classical helicase activity, but this is very weak in the absence of an assisting force. HELB binds specifically to human RPA, which enhances its ATPase and ssDNA translocase activities but inhibits DNA unwinding. Direct observation of HELB on RPA nucleoprotein filaments shows that translocating HELB concomitantly clears RPA from ssDNA. This activity, which can allow other proteins access to ssDNA intermediates despite their shielding by RPA, may underpin the diverse roles of HELB in cellular DNA transactions.[Significance] Single-stranded DNA (ssDNA) is a key intermediate in many cellular DNA transactions, including DNA replication, repair, and recombination. Nascent ssDNA is rapidly bound by the Replication Protein A (RPA) complex, forming a nucleoprotein filament that both stabilizes ssDNA and mediates downstream processing events. Paradoxically, however, the very high affinity of RPA for ssDNA may block the recruitment of further factors. In this work, we show that RPA–ssDNA nucleoprotein filaments are specifically targeted by the human HELB helicase. Recruitment of HELB by RPA–ssDNA activates HELB translocation activity, leading to processive removal of upstream RPA complexes. This RPA clearance activity may underpin the diverse roles of HELB in replication and recombination.Work in the laboratory of M.S.D. was supported by an Elizabeth Blackwell Early Career Fellowship from the University of Bristol (to O.J.W.) and Wellcome Trust Investigator Grant 100401/Z/12/Z (to M.S.D.). Work in the laboratory of E.A. was supported by NIH Grants GM130746 (to E.A.) and GM133967 (to E.A.). F.M.-H. acknowledges support from the European Research Council under European Union Horizon 2020 Research and Innovation Program Grant Agreement 681299. Work in the laboratory of F.M.-H. was also supported by Spanish Ministry of Science and Innovation Grants BFU2017-83794-P (AEI/FEDER, UE; to F.M.-H.) and PID2020-112998GB-100 (AEI/10.13039/501100011033; to F.M.-H.) and Comunidad de Madrid Grants Tec4-Bio–S2018/NMT-4443 (to F.M.-H.) and NanoBioCancer–Y2018/BIO-4747 (to F.M.-H.)

Gravitational Mesoscopic Constraints in Cosmological Dark Matter Halos

We present an analysis of the behaviour of the `coarse-grained' (`mesoscopic') rank partitioning of the mean energy of collections of particles composing virialized dark matter halos in a Lambda-CDM cosmological simulation. We find evidence that rank preservation depends on halo mass, in the sense that more massive halos show more rank preservation than less massive ones. We find that the most massive halos obey Arnold's theorem (on the ordering of the characteristic frequencies of the system) more frequently than less massive halos. This method may be useful to evaluate the coarse-graining level (minimum number of particles per energy cell) necessary to reasonably measure signatures of `mesoscopic' rank orderings in a gravitational system.Comment: LaTeX, 15 pages, 3 figures. Accepted for publication in Celestial Mechanics and Dynamical Astronomy Journa

Simulaçao numérica da dispersao de poluentes emitidos por incendios no verao de 2003 na Peninsula Ibérica

Ponencia presentada en: XXIX Jornadas Científicas de la AME y el VII Encuentro Hispano Luso de Meteorología celebrado en Pamplona, del 24 al 26 de abril de 2006.[PT]O objectivo deste trabalho é utilizar o modelo CATT-B-RAMS (Coupled Aerosol and Tracers Transport model to the Brazilian Regional Atmospheric Modeling System) para estudar o transporte atmosférico do monóxido de carbono (CO) e material particulado (PM2.5) emitidos por queimadas que ocorreram durante o Verão de 2003 na Península Ibérica, no período de 31 de Julho a 3 de Agosto com a situação sinóptica actuante na exportação destes poluentes. As simulações numéricas foram feitas com a assimilação dos dados de fogos derivados a partir de medidas do MODIS/AQUA para a Europa para implementar a posição da emissão. Para a caracterização do estado inicial do modelo foram usadas as análises do modelo global AVN/NCEP (Aviation run of the National Center for Environmental Prediction Global Spectral Model) com grade de resolução de 80 km. Os resultados indicam que o modelo de mesoescala pode ser uma boa ferramenta para descrever a circulação atmosférica reproduzindo as principais características da situação sinóptica e o entendimento e avaliação deste impacto, passam necessariamente, pela junção de estudos observacionais e modelação numérica gerando modelos complexos que descrevam as inter-relações biosfera-atmosfera, caracterizando um estudo multidisciplinar.[EN]This study presents the transport of gases and particulate emitted by large scale vegetation fires that took place in the Iberian Peninsula in summer of 2003. This study is carried out through a numerical simulation using a highresolution configuration of the 3-D transport model CATT-BRAMS (Coupled Aerosol and Tracer Transport to the Brazilian developments on the Regional Atmospheric Modeling System) coupled to a biomass burning emission model. The sources emission from biomass burning for carbon monoxide (CO) and particulate material (PM2.5) are defined using MODIS/TERRA fire product and local observations. The initial and lateral boundary conditions necessary to drive model were provided by the Aviation run of the National Center for Environmental Prediction Global Spectral Model (AVN/NCEP). The coarse grid resolution was defined with 80 km grid spacing. The results show that the mesoscale model can be used as a useful tool to describe the atmospheric circulation reproducing the main characteristics of the synoptic situation and how it controls the pollution transport