McAlister Victor J, Christie Gail E
Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, VA 23298-0678, USA.
J Bacteriol. 2009 Jul;191(14):4513-21. doi: 10.1128/JB.00193-09. Epub 2009 May 15.
The Serratia marcescens NucC protein is structurally and functionally homologous to the P2 Ogr family of eubacterial zinc finger transcription factors required for late gene expression in P2- and P4-related bacteriophages. These activators exhibit site-specific binding to a conserved DNA sequence, TGT-N(3)-R-N(4)-Y-N(3)-aCA, that is located upstream of NucC-dependent S. marcescens promoters and the late promoters of P2-related phages. In this report we describe the interactions of NucC with the P2 FETUD late operon promoter P(F). NucC is shown to bind P(F) as a tetramer and to make 12 symmetrical contacts to the DNA phosphodiester backbone. The backbone contacts are centered on the TGT-N(3)-R-N(4)-Y-N(3)-aCA motif. Major groove base contacts can be seen at most positions within the approximately 24-bp binding site. Minor groove contacts map to adjacent positions in the downstream half of the binding site, which corresponds to the area in which the DNA also appears to be bent by NucC binding. NucC binding provides a new example of protein-DNA interaction that is strikingly different from the DNA binding demonstrated for eukaryotic zinc-finger transcription factors.
粘质沙雷氏菌NucC蛋白在结构和功能上与P2和P4相关噬菌体中晚期基因表达所需的真细菌锌指转录因子的P2 Ogr家族同源。这些激活剂与保守的DNA序列TGT-N(3)-R-N(4)-Y-N(3)-aCA表现出位点特异性结合,该序列位于NucC依赖的粘质沙雷氏菌启动子和P2相关噬菌体的晚期启动子上游。在本报告中,我们描述了NucC与P2 FETUD晚期操纵子启动子P(F)的相互作用。结果表明,NucC以四聚体形式结合P(F),并与DNA磷酸二酯主链形成12个对称接触。主链接触以TGT-N(3)-R-N(4)-Y-N(3)-aCA基序为中心。在大约24 bp的结合位点内的大多数位置都可以看到大沟碱基接触。小沟接触映射到结合位点下游一半的相邻位置,这对应于DNA似乎也因NucC结合而弯曲的区域。NucC结合提供了一个蛋白质-DNA相互作用的新例子,它与真核锌指转录因子所展示的DNA结合显著不同。