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Crl; Transcriptome comparison of the crl mutant and CDT1-RNAi line

ABSTRACT: au13-04_cdtbis; cdt1_bis crl mutants and CDT1-RNAi lines have very similar macroscopic phenotypes as well as identical defects in plastid division and biogenesis. Our goal wwas to determine how much of these similarities originated from similar alterations of gene expression. Plantlets of crl mutant and CDT1-RNAi lines were grown in vitro on MS1/2 medium for 14 days. CDT1-RNAi lines were compared to the corresponding wild-type (Ws), whereas crl mutants were compared to their wild-type siblings and are in the Col0 ecotype.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE48465 | GEO | 2013/09/01

SECONDARY ACCESSION(S): PRJNA210593

REPOSITORIES: GEO

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cdt1_bis; Transcriptome comparison of the crl mutant and CDT1-RNAi line

Project description:au13-04_cdtbis; cdt1_bis crl mutants and CDT1-RNAi lines have very similar macroscopic phenotypes as well as identical defects in plastid division and biogenesis. Our goal wwas to determine how much of these similarities originated from similar alterations of gene expression. Plantlets of crl mutant and CDT1-RNAi lines were grown in vitro on MS1/2 medium for 14 days. CDT1-RNAi lines were compared to the corresponding wild-type (Ws), whereas crl mutants were compared to their wild-type siblings and are in the Col0 ecotype. 4 dye-swap - normal vs rnai mutant comparaison, normal vs transgenic comparaison

2013-09-01 | E-GEOD-48465 | biostudies-arrayexpress

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Project description:The origin recognition complex (ORC) nucleates DNA replication initiation in eukaryotic cells. This six-protein complex binds replication origin DNA, recruits other initiation factors and facilitates loading of the DNA helicase. Studying the function of individual ORC subunits during pre-RC formation has been hampered by the requirement of most subunits for DNA binding. In this study, we investigate the function of the S. cerevisiae Orc6 subunit, the only subunit not required for DNA binding. In vivo, depletion of Orc6 inhibits pre-replicative complex (pre-RC) assembly and maintenance. In vitro, ORC lacking Orc6 fails to interact with Cdt1 and to load the Mcm2-7 helicase onto origin DNA. We demonstrate that two regions of Orc6 bind Cdt1 directly and that the extreme C-terminus of Orc6 (Orc6-CTD) interacts tightly with the remaining five ORC subunits. Replacing Orc6 with a fusion protein linking Cdt1 to the Orc6-CTD results in an ORC complex that loads Mcm2-7 onto DNA. Interestingly, this complex can only perform a single round of Mcm2-7 loading, suggesting that a dynamic association of Cdt1 with ORC is required for multiple rounds of pre-RC assembly. Keywords: ChIP-chip

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CSN5i-3 induces degradation of selective CRL substrate receptors and accumulation of CRL substrates

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Transcription profiling of stationary phase E.coli wild type and crl::cat mutant strains to identify Crl-regulated genes

Project description:We wished to identify Crl-regulated genes in stationary phase in E.coli, and whether those overlap with the previously identified regulon of RpoS. Therefore wildtype E.coli (MC4100) and its isogenic crl::cat mutant were grown at 30oC. Total RNA was extracted at on OD (578nm) of 4 (during entry into stationary phase) and then the analysis proceeded as described in detail in the protocoles. The experiment was repeated three times and the results from those experiments are presented here.

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