Seed-specific transcription factor HSFA9 links embryogenesis and photomorphogenesis

Poster presentado en la XIII Reunión de Biología Molecular de Plantas. Oviedo 22-24 de junio de 2016 Posler 14/ $IV P14HSFA9, a seed-specific factor, enhances the expression of light receptors and genes required for chlorophyll biosynthesis before germinating seeds are illuminated. HSFA9 subsequently augments phytochrome-mediated responses and stimulates seedling greening.This work has been funded by FEDER (European Regional Development Fund) and by “Secretaría de Estado de Investigación, Desarrollo e Innovación” (projects BIO2011-23440 and BIO2014-52303-R). Additional funds were obtained from “Junta de Andalucía” (Group BIO148).Peer reviewe

Hahb-4, a homeobox-leucine zipper gene potentially involved in abscisic acid-dependent responses to water stress in sunflower

The nucleotide sequence data reported will appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession numbers AF339748 and AF339749.Homeodomain-leucine zipper proteins constitute a family of transcription factors found only in plants. We have characterized a full-length cDNA encoding the sunflower homeobox-leucine zipper protein Hahb-4 (Helianthus annuus homeo box-4). The complete cDNA is 674 base pairs long and contains an open reading frame of 177 amino acids that belongs to the Hd-Zip I subfamily. Northern blot and RNAse A protection analysis show that the expression of Hahb-4 is rapidly, strongly and reversibly induced by water deficit in whole seedlings, roots, stems and leaves. A similar fast induction of Hahb-4 expression is observed when seedlings are subjected to a treatment with the hormone abscisic acid (ABA). Nuclei prepared from seedlings treated with ABA or subjected to water stress show a significant increase of protein(s) that specifically bind the sequence 5′-CAAT(A/T)ATTG-3′, recognized in vitro by Hahb-4, suggesting that an active protein is synthesized in response to these treatments. The promoter region of the Hahb-4 gene contains sequences that fit the consensus for a G-box present in some ABA responsive elements (ABREs). We propose that Hahb-4 may function in signalling cascade(s) that control(s) a subset of the ABAmediated responses of sunflower to water stress.This work was supported by grants from CONICET, ANPCyT, Fundación Antorchas (Argentina) and Universidad Nacional del Litoral. R.L.C. and D.H.G. are members of CONICET; G.M.G. is a fellow of the same Institution. The laboratory work of J.J. and C.A. was also supported by grants BIO99-794 (from the Spanish Comisión Interministerial de Ciencia y Tecnología) and CVI 148 (from Plan Andaluz de Investigación).Peer reviewe

Seed-specific transcription factor HSFA9 links late embryogenesis and early photomorphogenesis

HSFA9 is a seed-specific transcription factor that in sunflower (Helianthus annuus) is involved in desiccation tolerance and longevity. Here we show that the constitutive overexpression of HSFA9 in tobacco (Nicotiana tabacum) seedlings attenuated hypocotyl growth under darkness and accelerated the initial photosynthetic development. Plants overexpressing HSFA9 increased accumulation of carotenoids, chlorophyllide, and chlorophyll, and displayed earlier unfolding of the cotyledons. HSFA9 enhanced phytochrome-dependent light responses, as shown by an intensified hypocotyl length reduction after treatments with continuous far-red or red light. This observation indicated the involvement of at least two phytochromes: PHYA and PHYB. Reduced hypocotyl length under darkness did not depend on phytochrome photo-activation; this was inferred from the lack of effect observed using far-red light pulses applied before the dark treatment. HSFA9 increased the expression of genes that activate photomorphogenesis, including PHYA, PHYB, and HY5. HSFA9 might directly upregulate PHYA and indirectly affect PHYB transcription, as suggested by transient expression assays. Converse effects on gene expression, greening, and cotyledon unfolding were observed using a dominant-negative form of HSFA9, which was overexpressed under a seed-specific promoter. This work uncovers a novel transcriptional link, through HSFA9, between seed maturation and early photomorphogenesis. In all, our data suggest that HSFA9 enhances photomorphogenesis via early transcriptional effects that start in seeds under darkness

The HaDREB2 transcription factor enhances basal thermotolerance and longevity of seeds through functional interaction with HaHSFA9

<p>Abstract</p> <p>Background</p> <p>Transcription factor HaDREB2 was identified in sunflower (<it>Helianthus annuus </it>L.) as a drought-responsive element-binding factor 2 (DREB2) with unique properties. HaDREB2 and the sunflower Heat Shock Factor A9 (HaHSFA9) co-activated the <it>Hahsp17.6G1 </it>promoter in sunflower embryos. Both factors could be involved in transcriptional co-activation of additional small heat stress protein (sHSP) promoters, and thus contribute to the HaHSFA9-mediated enhancement of longevity and basal thermotolerance of seeds.</p> <p>Results</p> <p>We found that overexpression of HaDREB2 in seeds did not enhance longevity. This was deduced from assays of basal thermotolerance and controlled seed-deterioration, which were performed with transgenic tobacco. Furthermore, the constitutive overexpression of HaDREB2 did not increase thermotolerance in seedlings or result in the accumulation of HSPs at normal growth temperatures. In contrast, when HaDREB2 and HaHSFA9 were conjointly overexpressed in seeds, we observed positive effects on seed longevity, beyond those observed with overexpression of HaHSFA9 alone. Such additional effects are accompanied by a subtle enhancement of the accumulation of subsets of sHSPs belonging to the CI and CII cytosolic classes.</p> <p>Conclusion</p> <p>Our results reveal the functional interdependency of HaDREB2 and HaHSFA9 in seeds. HaDREB2 differs from other previously characterized DREB2 factors in plants in terms of its unique functional interaction with the seed-specific HaHSFA9 factor. No functional interaction between HaDREB2 and HaHSFA9 was observed when both factors were conjointly overexpressed in vegetative tissues. We therefore suggest that additional, seed-specific factors, or protein modifications, could be required for the functional interaction between HaDREB2 and HaHSFA9.</p

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Developmental and environmental concurrent expression of sunflower dry-seed-stored low-molecular-weight heat-shock protein and Lea mRNAs

We have cloned and sequenced three different cDNAs from sunflower seed-stored mRNA. Sequence similarities and response to heat-shock identified one of the cDNAs as a low-molecular-weight heat-shock protein (lmw-HSP). The other two clones showed significant sequence similarity to the cotton and carrot late-embryogenesis-abundant (Lea) proteins D-113 and Emb-1, respectively. The three cDNAs showed similar expression patterns during zygotic embryo development, as well as in vegetative tissues of 3-day-old seedlings in response to stress. Maximal accumulation of all three mRNAs was detected in dry seeds and during embryo mid-maturation stage, in the absence of exogenous stress. In seedlings, mRNAs accumulated to lower levels in response to osmotic stress and exogenous abscisic acid (ABA) treatments. A differential time course of response to osmotic stress was observed: lmw-HSP mRNA accumulation was induced earlier than that of Lea mRNAs. The coordinate accumulation of Lea and lmw-HSP transcripts during embryo development and in response to stress and ABA suggests the existence of common regulatory elements for Lea and lmw-HSP genes, and supports the notion that HSPs might have alternative functions in the plant cell.Was the recipient of a postdoctoral ('reincorporacidn') fellowship from the Ministerio de Educaci6n y Ciencia. This work was supported by grant BIO90-0759 from CICYT.Peer reviewe

Expression of sunflower low-molecular-weight heat-shock proteins during embryogenesis and persistence after germination: localization and possible functional implications

We isolated and sequenced Ha hsp17.9, a DNA complementary (cDNA) of dry-seed stored mRNA that encodes a low-molecular-weight heat-shock protein (LMW HSP). Sequence analysis identified Ha hspl7.9, and the previously reported Ha hspl7.6, as cDNAs encoding proteins (HSP17.6 and H S P 17.9) which belong to different families of cytoplasmic LMW H S Ps. Using specific antibodies we observed differential expression of both proteins during zygotic embryogenesis under controlled environment, and a remarkable persistence of these LMW HSPs during germination. Immuno-blot analysis of HSP17.9 proteins in two-dimensional gels revealed that the polypeptides expressed in embryos were indistinguishable from LMW HSPs expressed in vegetative tissues in response to water deficit; but they appeared different from homologeous proteins expressed in response to thermal-stress. Tissue-print immunolocalization experiments showed that HSP17.9 and HSP17.6 were homogeneously distributed in every tissue of desiccation-tolerant dry seeds and young seedlings under non-stress conditions. These results demonstrate developmental regulation of specific, cytoplasmic, plant LMW HSPs, suggesting also their involvement in water-stress tolerance.Was supported by a Ph.D. fellowship from the Spanish 'Ministerio de Educaci6n y Ciencia'. CA was a recipient of a post-doctoral ('reincorporaci6n') fellowship from Spanish CICYT. This work was funded through grants BIO90-0759 and BIO93-0023 from CICYT.Peer reviewe

Detection of high incidence of K-ras oncogenes during human colon tumorigenesis

RNAse A mismatch cleavage analysis of 66 primary human colon tumours reveals a high incidence of K-ras genes with mutations at position 12. No apparent correlation was found between the presence of mutant oncogenes and the degree of invasiveness of the tumours but evidence for ras mutational activation in premalignant tissue was obtained.This work was supported by an NIH grant awarded by tbe Nalional Cancer Institute and by a grant from Toyo Jozo Ltd.Peer reviewe

Constitutive expression of small heat shock proteins in vegetative tissues of the resurrection plant craterostigma plantagineum

Using antibodies raised against two sunflower small heat shock proteins (sHSPs), we have detected immunologically related proteins in unstressed vegetative tissues from the resurrection plant Craterostigma plantagineum. In whole plants, further accumulation of these polypeptides was induced by heat-shock or water-stress. In desiccation-intolerant Craterostigma callus tissue, we failed to detect sHSP-related polypeptides, but their expression, and the concurrent acquisition of desiccation tolerance was induced by exogenous abscisic acid (ABA) treatment. In untressed plants, the cross-reacting polypeptides were abundant in the roots and lower part of the shoots, where they showed homogeneous tissue-distributions. This constitutive expression is novel for vegetative tissues of higher plants, and resembles the expression patterns of sHSPs in desiccation-tolerant zygotic embryos and germinating seeds.Was supported by a Human Capital Mobility EU fellowship. CA was a recipient of a post-doctoral ('reincorporación') contract from Spanish CICYT. Collaboration between the Cologne and Seville groups was inspired in meetings organized under the Interdrought'EU concerted action (AIR3-CT92-0742). Part of this work was funded through grant BIO93-0023, awarded to J.J.Peer reviewe