Zhang S, Burkett T J, Yamashita I, Garfinkel D J
NCI-Frederick Cancer Research and Development Center, ABL-Basic Research Program, Maryland 21702-1201, USA.
Mol Cell Biol. 1997 Aug;17(8):4718-29. doi: 10.1128/MCB.17.8.4718.
SPT23 was isolated as a dosage-dependent suppressor of Ty-induced mutations in Saccharomyces cerevisiae. SPT23 shows considerable sequence homology with MGA2, a gene identified as a dosage-dependent suppressor of a snf2-imposed block on STA1 transcription in S. cerevisiae var. diastaticus. Although single mutations in either of these genes have only modest effects on cell growth, spt23 mga2 double mutants are inviable. Unlike SPT23, multicopy expression of a truncated form of MGA2 suppresses a narrow subset of Ty-induced mutations. SPT23/MGA2 and the SNF/SWI genes affect transcription of certain target genes in similar ways. Spt23p appears to be a rate-limiting component required for functional HIS4 expression of his4-912delta, a promoter insertion mutation induced by the Ty1-912 long terminal repeat. Furthermore, both Spt23p and Mga2p can activate transcription when fused to the Gal4p DNA-binding domain, as previously observed with Snf2p and Snf5p. A 50-amino-acid region in the N terminus of the predicted Spt23p protein is necessary and sufficient for the transactivation and necessary for suppression of Ty1-induced mutations and the essential function of Spt23p. Cell fractionation and cytological experiments suggest that Spt23p is associated with the nucleus. Our results suggest that SPT23/MGA2 affects transcription of a subset of genes in yeast, perhaps by changing chromatin accessibility.
SPT23是作为酿酒酵母中Ty诱导突变的剂量依赖性抑制因子而分离得到的。SPT23与MGA2具有相当程度的序列同源性,MGA2是一个被鉴定为酿酒酵母变种diastaticus中snf2对STA1转录施加阻断的剂量依赖性抑制因子的基因。尽管这两个基因中的任何一个发生单突变对细胞生长只有适度影响,但spt23 mga2双突变体是无法存活的。与SPT23不同,截短形式的MGA2的多拷贝表达只能抑制一小部分Ty诱导的突变。SPT23/MGA2和SNF/SWI基因以相似的方式影响某些靶基因的转录。Spt23p似乎是his4-912delta功能性HIS4表达所必需的限速成分,his4-912delta是由Ty1-912长末端重复序列诱导的启动子插入突变。此外,正如之前在Snf2p和Snf5p中观察到的那样,当与Gal4p DNA结合结构域融合时,Spt23p和Mga2p都可以激活转录。预测的Spt23p蛋白N端的一个50个氨基酸的区域对于反式激活是必要且充分的,对于抑制Ty1诱导的突变和Spt23p的基本功能也是必要的。细胞分级分离和细胞学实验表明Spt23p与细胞核相关。我们的结果表明SPT23/MGA2可能通过改变染色质可及性来影响酵母中一部分基因的转录。