Cell differentiation by interaction of two HMG-box proteins: Mat1-Mc activates M cell-specific genes in S-pombe by recruiting the ubiquitous transcription factor Ste11 to weak binding sites

被引:44
作者
Kjaerulff, S
Dooijes, D
Clevers, H
Nielsen, O
机构
[1] UNIV COPENHAGEN, INST MOL BIOL, DEPT GENET, DK-1353 COPENHAGEN K, DENMARK
[2] UNIV UTRECHT HOSP, DEPT IMMUNOL, NL-3508 GA UTRECHT, NETHERLANDS
关键词
differentiation; fission yeast; HMG-box proteins; sex determination; transcriptional activation;
D O I
10.1093/emboj/16.13.4021
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The Schizosaccharomyces pombe mfm1 gene is expressed in an M cell-specific fashion, This regulation requires two HMG-box proteins: the ubiquitous Ste11 transcription factor and the M cell-controlling protein Mat1-Mc. Here we report that the mfm1 promoter contains a single, weak Ste11-binding site (a so-called TR-box) that can confer M-specificity on a heterologous promoter when present in eight copies, In vitro, both Mat1-Mc and Ste11 can bind this box with approximately the same affinity, The Mat1-Mc protein caused a dramatic increase in the DNA-binding of Ste11 to this box, under conditions where we could not detect Mat1-Mc in the resulting protein-DNA complex. When we changed a single base in the mfm1 TR-box, such that it resembled those boxes found in ubiquitously expressed genes, Ste11 binding was enhanced, and in vivo the mfm1 gene also became expressed in P cells where Mat1-Mc is absent, These findings suggest that M-specificity results from Mat1-Mc-mediated Ste11 binding to weak TR-boxes, We have also defined a novel moth (termed M-box), adjacent to the mfm1 TR-box, to which Mat1-Mc binds strongly, A DNA fragment containing both the TR- and the M-box allowed the formation of a complex containing both Ste11 and Mat1-Mc, A single copy of this fragment was sufficient to activate a heterologous promoter in an M-specific fashion, suggesting that these two boxes act in a synergistic manner.
引用
收藏
页码:4021 / 4033
页数:13
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