Gailus-Durner V, Xie J, Chintamaneni C, Vershon A K
Waksman Institute, Rutgers University, Piscataway, New Jersey 08855-0759, USA.
Mol Cell Biol. 1996 Jun;16(6):2777-86. doi: 10.1128/MCB.16.6.2777.
The meiosis-specific gene HOP1, which encodes a component of the synaptonemal complex, is controlled through two regulatory elements, UASH and URS1H. Sites similar to URS1H have been identified in the promoter region of virtually every early meiosis-specific gene, as well as in many promoters of nonmeiotic genes, and it has been shown that the proteins that bind to this site function to regulate meiotic and nonmeiotic transcription. Sites similar to the UASH site have been found in a number of meiotic and nonmeiotic genes as well. Since it has been shown that UASH functions as an activator site in vegetative haploid cells, it seemed likely that the factors binding to this site regulate both meiotic and nonmeiotic transcription. We purified the factor binding to the UASH element of the HOP1 promoter. Sequence analysis identified the protein as Abf1 (autonomously replicating sequence-binding factor 1), a multifunctional protein involved in DNA replication, silencing, and transcriptional regulation. We show by mutational analysis of the UASH site, that positions outside of the proposed UASH consensus sequence (TNTGN[A/T]GT) are required for DNA binding in vitro and transcriptional activation in vivo. A new UASH consensus sequence derived from this mutational analysis closely matches a consensus Abf1 binding site. We also show that an Abf1 site from a nonmeiotic gene can replace the function of the UASH site in the HOP1 promoter. Taken together, these results show that Abf1 functions to regulate meiotic gene expression.
减数分裂特异性基因HOP1编码联会复合体的一个组成部分，它受两个调控元件UASH和URS1H的控制。几乎在每一个早期减数分裂特异性基因的启动子区域，以及许多非减数分裂基因的启动子中，都发现了与URS1H相似的位点，并且已经证明，与该位点结合的蛋白质具有调控减数分裂和非减数分裂转录的功能。在许多减数分裂和非减数分裂基因中也发现了与UASH位点相似的位点。由于已经证明UASH在营养单倍体细胞中作为激活位点发挥作用，所以与该位点结合的因子似乎有可能同时调控减数分裂和非减数分裂转录。我们纯化了与HOP1启动子的UASH元件结合的因子。序列分析确定该蛋白质为Abf1（自主复制序列结合因子1），这是一种参与DNA复制、沉默和转录调控的多功能蛋白质。我们通过对UASH位点的突变分析表明，在体外DNA结合和体内转录激活过程中，拟议的UASH共有序列（TNTGN[A/T]GT）之外的位置是必需的。从该突变分析得出的新的UASH共有序列与Abf1共有结合位点紧密匹配。我们还表明，来自非减数分裂基因的一个Abf1位点可以取代HOP1启动子中UASH位点的功能。综上所述，这些结果表明Abf1具有调控减数分裂基因表达的功能。
