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HAP5 Triple Mutant vs. Constans Mutant

ABSTRACT: Aims: To compare the transcriptional profiles of an AtHAP5 (AtNF-YC, AtCBF-C) triple mutant vs. a Constans mutant. Background Information on Mutants: AtHAP5 triple mutant created by cross between the following SALK TDNA inserts: SALK_034838, SALK_058903, SALK_032163; these correspond to At1G54830, At1G08970, At5G63470, respectively. co-SAIL TDNA insertion SAIL_24_H04 corresponds to AT5G15840. Tissue: Above-ground whole plants grown for 10 days, under 16 hr light 8 hrs dark at constant 22C. Tissue collected 14 hrs after lights on. RT-PCR analysis on test RNA confirmed FT levels reduced in mutants lines compared to wild-type.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE20258 | GEO | 2010/04/20

SECONDARY ACCESSION(S): PRJNA124221

REPOSITORIES: GEO

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HAP5 Triple Mutant vs. Constans Mutant

Project description:Aims: To compare the transcriptional profiles of an AtHAP5 (AtNF-YC, AtCBF-C) triple mutant vs. a Constans mutant. Background Information on Mutants: AtHAP5 triple mutant created by cross between the following SALK TDNA inserts: SALK_034838, SALK_058903, SALK_032163; these correspond to At1G54830, At1G08970, At5G63470, respectively. co-SAIL TDNA insertion SAIL_24_H04 corresponds to AT5G15840. Tissue: Above-ground whole plants grown for 10 days, under 16 hr light 8 hrs dark at constant 22C. Tissue collected 14 hrs after lights on. RT-PCR analysis on test RNA confirmed FT levels reduced in mutants lines compared to wild-type. 9 samples were used in this experiment.

2010-04-28 | E-GEOD-20258 | biostudies-arrayexpress

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Project description:Nuclear Factor Y (NF-Y) is a heterotrimeric transcription factor that binds CCAAT elements. The NF-Y trimer is composed of a Histone Fold Domain (HFD) dimer (NF-YB/NF-YC) and NF-YA, which confers DNA sequence specificity. NF-YA shares a conserved domain with the CONSTANS, CONSTANS-LIKE, TOC1 (CCT) proteins. We show that CONSTANS (CO/B-BOX PROTEIN1 BBX1), a master flowering regulator, forms a trimer with Arabidopsis thaliana NF-YB2/NF-YC3 to efficiently bind the CORE element of the FLOWERING LOCUS T promoter. Using saturation mutagenesis, electrophoretic mobility shift assays, and RNA-sequencing profiling of co, nf-yb, and nf-yc mutants, we identify CCACA elements as the core NF-CO binding site. CO physically interacts with the same HFD surface required for NF-YA association, as determined by mutations in NF-YB2 and NF-YC9, and tested in vitro and in vivo. The co-7 mutation in the CCT domain, corresponding to an NF-YA arginine directly involved in CCAAT recognition, abolishes NF-CO binding to DNA

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Project description:HAP5 Triple Mutant vs. Constans Mutant

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Characterization of miP1a/b (microProteins) overexpression plants, including a constans mutant (co-SAIL)

Project description:MicroProteins are short, single domain proteins that act by sequestering larger, multidomain proteins into non-functional complexes. MicroProteins have been identified in plants and animals, where they are mostly involved in the regulation of developmental processes. Here we show that two Arabidopsis thaliana microProteins, miP1a and miP1b, physically interact with CONSTANS (CO) a potent regulator of flowering time. The miP1a/b-type microProteins evolved in dicotyledonous plants and have an additional carboxy-terminal PF(V/L)FL motif. This motif enables miP1a/b microProteins to interact with TOPLESS/TOPLESS-RELATED (TPL/TPR) proteins. Interaction of CO with miP1a/b/TPL causes late flowering due to a failure in the induction of FLOWERING LOCUS T (FT) expression under inductive long day conditions. Both miP1a and miP1b are expressed in vascular tissue, where CO and FT are active. Genetically, miP1a/b act upstream of CO thus our findings unravel a novel layer of flowering time regulation via microProtein-inhibition. RNA-Seq transcriptome analysis of four biological samples were analysed with two technical replicates. Columbia wildtype plants Col-0, constans mutant plants co-SAIL, and two transgenic lines overexpressing a microProtein (miP1a and miP1b) were sequenced.

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