Low D, Blyn L
Department of Pathology, University of Utah School of Medicine, Salt Lake City 84132.
Rev Infect Dis. 1988 Jul-Aug;10 Suppl 2:S300-5. doi: 10.1093/cid/10.supplement_2.s300.
Uropathogenic Escherichia coli are major causative agents of cystitis and pyelonephritis. Most E. coli pyelonephritis isolates express pili encoded by the pyelonephritis-associated pili (pap) gene cluster. The pap DNA sequence encodes pilin monomers that are assembled into pili fibers; pap also encodes adhesins that recent results suggest might be located at the tips of the pilus fibers. The study presented here is a status report of work that has two major goals: to determine (1) if any Pap proteins associate with pili and (2) if Pap proteins not required for pili assembly affect levels of pili-cell surface expression. To address the first aim, antisera to pili were used to precipitate pili from detergent extracts containing 35S-labeled Pap proteins. The results suggested that a protein of 16-kilodaltons apparent molecular mass associated with pili. Other interpretations of the data are discussed. The second aim was addressed by constructing E. coli strains that contained different pap regions. With the use of electron microscopy and a pili ELISA, it was found that E. coli containing a 6.5-kilobase-pair region of pap expressed low levels of pili, but no P-adhesin was detected. Transformation of this E. coli strain with a plasmid containing an additional 3.5-kilobase-pair pap DNA sequence resulted in an eightfold increase in pili expression as well as expression of P adhesin. These results indicated that pili expression was affected by Pap proteins not required for pili assembly.
尿路致病性大肠杆菌是膀胱炎和肾盂肾炎的主要病原体。大多数大肠杆菌肾盂肾炎分离株表达由肾盂肾炎相关菌毛(pap)基因簇编码的菌毛。pap DNA序列编码组装成菌毛纤维的菌毛蛋白单体;pap还编码黏附素,最近的研究结果表明这些黏附素可能位于菌毛纤维的尖端。本文介绍的研究是一项工作进展报告,有两个主要目标:确定(1)是否有任何Pap蛋白与菌毛相关,以及(2)菌毛组装不需要的Pap蛋白是否会影响菌毛在细胞表面的表达水平。为实现第一个目标,使用针对菌毛的抗血清从含有35S标记的Pap蛋白的去污剂提取物中沉淀菌毛。结果表明,一种表观分子量为16千道尔顿的蛋白质与菌毛相关。文中还讨论了对这些数据的其他解释。通过构建含有不同pap区域的大肠杆菌菌株来实现第二个目标。利用电子显微镜和菌毛酶联免疫吸附测定法发现,含有6.5千碱基对pap区域的大肠杆菌表达的菌毛水平较低,但未检测到P黏附素。用含有额外3.5千碱基对pap DNA序列的质粒转化该大肠杆菌菌株,导致菌毛表达增加了八倍,同时也表达了P黏附素。这些结果表明,菌毛组装不需要的Pap蛋白会影响菌毛的表达。