Schandel K A, Maneewannakul S, Ippen-Ihler K, Webster R E
J Bacteriol. 1987 Jul;169(7):3151-9. doi: 10.1128/jb.169.7.3151-3159.1987.
An F lac pro mutant which was temperature sensitive for infection by the filamentous bacteriophage f1 but resistant to the F-specific icosahedral RNA phage f2 was isolated. Cells carrying the F' mutation failed to elaborate F pili at all temperatures. Mutant cells were able to pair with recipient cells during bacterial conjugation, but transfer of conjugal DNA occurred at a greatly reduced frequency. Complementation analyses showed the F' mutation to be in the traC gene. When a plasmid carrying traC was introduced into hosts harboring the F' mutation, phage sensitivity, the ability to elaborate F pili, and conjugation efficiency were restored. The mutation was named traC1044. The F lac pro traC1044 mutant appears to be unique among traC mutants in retaining host sensitivity to the filamentous phage f1 in the absence of expression of extended F pili. Phage f1 attachment sites appeared to be present at the cell surface in traC1044 mutants. The reduced accessibility of these sites may account for the reduced efficiency of phage f1 infection of traC1044 hosts, although the possibility that a defect was present in the receptor site itself was not eliminated. Membranes of hosts carrying the F' mutation contained a full complement of mature F-pilin subunits, so the product of traC is presumably required for pilus assembly but not for pilin processing. This, together with the deficiency in conjugal DNA transfer, suggests that traC may be part of a membrane-spanning tra protein complex responsible for pilus assembly and disassembly and conjugal DNA transmission.
分离出一种F lac pro突变体,它对丝状噬菌体f1的感染具有温度敏感性,但对F特异性二十面体RNA噬菌体f2具有抗性。携带F'突变的细胞在所有温度下都不能形成F菌毛。突变细胞在细菌接合过程中能够与受体细胞配对,但接合DNA的转移频率大大降低。互补分析表明F'突变位于traC基因中。当将携带traC的质粒导入含有F'突变的宿主中时,噬菌体敏感性、形成F菌毛的能力和接合效率得以恢复。该突变被命名为traC1044。F lac pro traC1044突变体在traC突变体中似乎是独特的,即在没有延长F菌毛表达的情况下仍保留宿主对丝状噬菌体f1的敏感性。在traC1044突变体中,噬菌体f1附着位点似乎存在于细胞表面。这些位点可及性的降低可能解释了traC1044宿主中噬菌体f1感染效率的降低,尽管受体位点本身存在缺陷的可能性并未排除。携带F'突变的宿主细胞膜含有完整的成熟F菌毛蛋白亚基,因此推测traC的产物是菌毛组装所必需的,但不是菌毛蛋白加工所必需的。这与接合DNA转移的缺陷一起表明,traC可能是负责菌毛组装和解体以及接合DNA传递的跨膜tra蛋白复合物的一部分。