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  • Sgf73, a subunit of SAGA complex, is required for the assembly of RITS complex in fission yeast.

Sgf73, a subunit of SAGA complex, is required for the assembly of RITS complex in fission yeast.

Scientific reports (2015-10-08)
Xiaolong Deng, Huan Zhou, Guiping Zhang, Wenchao Wang, Langyong Mao, Xing Zhou, Yao Yu, Hong Lu
ABSTRACT

RNA interference (RNAi) is a widespread gene-silencing mechanism and is required for heterochromatin assembly in a variety of organisms. The RNA-induced transcriptional silencing complex (RITS), composed of Ago1, Tas3 and Chp1, is a key component of RNAi machinery in fission yeast that connects short interference RNA (siRNA) and heterochromatin formation. However, the process by which RITS is assembled is not well understood. Here, we identified Sgf73, a subunit of the SAGA co-transcriptional complex, is required for pericentromeric heterochromatin silencing and the generation of siRNA. This novel role of Sgf73 is independent of enzymatic activities or structural integrity of SAGA. Instead, Sgf73 is physically associated with Ago1 and Chp1. The interactions among the subunits of the RITS, including those between Tas3 and Chp1, between Chp1 and Ago1, between Ago1 and Tas3, were all impaired by the deletion of sgf73(+). Consistently, the recruitment of Ago1 and Chp1 to the pericentromeric region was abolished in sgf73Δ cells. Our study unveils a moonlighting function of a SAGA subunit. It suggests Sgf73 is a novel factor that promotes assembly of RITS and RNAi-mediated heterochromatin formation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-dimethyl-Histone H3 (Lys9) Antibody, Upstate®, from rabbit
Sigma-Aldrich
EZ-Magna ChIP® A - Chromatin Immunoprecipitation Kit, Single day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using magnetic A beads. Control primers included.