Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function.
Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function.
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粪肠球菌接合质粒 pCF10 区域的克隆和表征，该区域编码性信息素结合功能。
DOI：
10.1128/jb.175.16.5253-5259.1993
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发表时间：
1993
影响因子：
3.2
通讯作者：
Dunny,GM
中科院分区：
文献类型：
--
作者：
Ruhfel,RE;Manias,DA;Dunny,GM
为了探究多肽性信息素诱导粪肠球菌质粒偶联表达的机制，采用生物学方法测定了携带偶联质粒pCF10的细胞与性信息素cCF10的结合能力。这些数据表明，pCF10赋予其宿主粪肠杆菌细胞特异性地从培养基中去除cCF10活性（显然是通过不可逆结合）的能力。编码这种能力的pCF10 DNA定位在一个3.4 kb的片段上，这个片段与负控制共轭传递函数的表达有关。该片段还编码了结合信息素抑制剂肽iCF10的能力。DNA测序显示了三个开放阅读框，分别为prgW（信息素响应基因W）、prgZ和prgY。推断出的prgW的产物类似于其他细菌和真核生物的调节蛋白，在假定的dna结合域内具有非常高的同一性。根据F.Y. an和D.B. Clewell（摘要）的研究，prgY基因实际上延伸到pCF10的邻近区域，并可以编码一种与TraB蛋白非常相似的蛋白，该蛋白被认为与携带信息素诱导溶血质粒pAD1的细胞关闭信息素cAD1的产生有关。将军见面。点。Soc。微生物学。1992,H70, 1992)。prgZ基因产物与革兰氏阳性和革兰氏阴性细菌中寡肽渗透酶（opp）操纵子编码的结合蛋白有显著相关性，并且与pad1编码的蛋白TraC高度相似，后者被认为介导性信息素cAD1的结合（K. Tanimoto， F. Y. An, and D. B. Clewell，提交发表）。将Tn5插入prgZ会破坏cCF10的结合能力。
In order to investigate the mechanism by which peptide sex pheromones induce expression of the conjugation functions of certain Enterococcus faecalis plasmids, a biological assay was developed to measure the ability of cells carrying the conjugative plasmid pCF10 to bind the sex pheromone cCF10. The data indicated that pCF10 endows its host E. faecalis cell with the ability to specifically remove (apparently by irreversible binding) cCF10 activity from culture medium. The pCF10 DNA encoding this ability was localized to a 3.4-kb segment within a region involved in negative control of expression of conjugal transfer functions. This segment also encoded ability to bind the pheromone inhibitor peptide iCF10. DNA sequencing revealed three open reading frames, which have been denoted prgW (pheromone responsive gene W), prgZ, and prgY. The deduced product of prgW resembled regulatory proteins from other bacteria and eucaryotes, with a very high degree of identity within a putative DNA-binding domain. The prgY gene actually extended into an adjacent region of pCF10 and could encode a protein with significant similarity to a protein called TraB, believed to be involved in shutdown of pheromone cAD1 production by cells carrying the pheromone-inducible hemolysin plasmid pAD1, according to F.Y. An and D.B. Clewell (Abstr. Gen. Meet. Am. Soc. Microbiol. 1992, H70, 1992). The prgZ gene product showed significant relatedness to binding proteins encoded by oligopeptide permease (opp) operons in gram-positive and gram-negative bacteria and is highly similar to a pAD1-encoded protein, TraC, which is believed to mediate sex pheromone cAD1 binding (K. Tanimoto, F. Y. An, and D. B. Clewell, submitted for publication). A Tn5 insertion into prgZ abolished cCF10 binding ability.