原创力文档reverse genetic analysis of the yeast rsc chromatin remodeler reveals a role for rsc3 and snf5 homolog 1 in ploidy maintenance反向遗传分析酵母rsc的染色质重塑揭示角色rsc3和snf5同族体1倍性维护.pdfVIP
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reverse genetic analysis of the yeast rsc chromatin remodeler reveals a role for rsc3 and snf5 homolog 1 in ploidy maintenance反向遗传分析酵母rsc的染色质重塑揭示角色rsc3和snf5同族体1倍性维护
Reverse Genetic Analysis of the Yeast RSC
Chromatin Remodeler Reveals a Role for RSC3
and SNF5 Homolog 1 in Ploidy Maintenance
*
Coen Campsteijn, Anne-Marie J. Wijnands-Collin, Colin Logie
Molecular Biology Department, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen, Netherlands
The yeast ‘‘remodels the structure of chromatin’’ (RSC) complex is a multi-subunit ‘‘switching deficient/sucrose non-
fermenting’’ type ATP-dependent nucleosome remodeler, with human counterparts that are well-established tumor
suppressors. Using temperature-inducible degron fusions of all the essential RSC subunits, we set out to map RSC
requirement as a function of the mitotic cell cycle. We found that RSC executes essential functions during G1, G2, and
mitosis. Remarkably, we observed a doubling of chromosome complements when degron alleles of the RSC subunit
SFH1, the yeast hSNF5 tumor suppressor ortholog, and RSC3 were combined. The requirement for simultaneous
deregulation of SFH1 and RSC3 to induce these ploidy shifts was eliminated by knockout of the S-phase cyclin CLB5 and
by transient depletion of replication origin licensing factor Cdc6p. Further, combination of the degron alleles of SFH1
and RSC3, with deletion alleles of each of the nine Cdc28/Cdk1-associated cyclins, revealed a strong and specific
genetic interaction between the S-phase cyclin genes CLB5 and RSC3, indicating a role for Rsc3p in proper S-phase
regulation. Taken together, our results implicate RSC in regulation of the G1/S-phase transition and establish a hitherto
unanticipated role for RSC-mediated chromatin remodeling in ploidy maintenance.
Citation: Campsteijn C, Wijnands-Collin AMJ, Logie C (2007) Reverse genetic analysis of the yeast RSC chromatin remodeler reveals a role for RSC3 and SNF5 homolog 1 in
ploidy maintena
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