Joslin Stephanie N, Pybus Christine, Labandeira-Rey Maria, Evans Amanda S, Attia Ahmed S, Brautigam Chad A, Hansen Eric J
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Infect Immun. 2015 Jan;83(1):146-60. doi: 10.1128/IAI.02486-14. Epub 2014 Oct 13.
There are a paucity of data concerning gene products that could contribute to the ability of Moraxella catarrhalis to colonize the human nasopharynx. Inactivation of a gene (mesR) encoding a predicted response regulator of a two-component signal transduction system in M. catarrhalis yielded a mutant unable to grow in liquid media. This mesR mutant also exhibited increased sensitivity to certain stressors, including polymyxin B, SDS, and hydrogen peroxide. Inactivation of the gene (mesS) encoding the predicted cognate sensor (histidine) kinase yielded a mutant with the same inability to grow in liquid media as the mesR mutant. DNA microarray and real-time reverse transcriptase PCR analyses indicated that several genes previously shown to be involved in the ability of M. catarrhalis to persist in the chinchilla nasopharynx were upregulated in the mesR mutant. Two other open reading frames upregulated in the mesR mutant were shown to encode small proteins (LipA and LipB) that had amino acid sequence homology to bacterial adhesins and structural homology to bacterial lysozyme inhibitors. Inactivation of both lipA and lipB did not affect the ability of M. catarrhalis O35E to attach to a human bronchial epithelial cell line in vitro. Purified recombinant LipA and LipB fusion proteins were each shown to inhibit human lysozyme activity in vitro and in saliva. A lipA lipB deletion mutant was more sensitive than the wild-type parent strain to killing by human lysozyme in the presence of human apolactoferrin. This is the first report of the production of lysozyme inhibitors by M. catarrhalis.
关于可能有助于卡他莫拉菌在人类鼻咽部定殖的基因产物的数据很少。对卡他莫拉菌中编码双组分信号转导系统预测反应调节因子的一个基因(mesR)进行失活,产生了一个无法在液体培养基中生长的突变体。这个mesR突变体对某些应激源,包括多粘菌素B、十二烷基硫酸钠和过氧化氢,也表现出更高的敏感性。对编码预测的同源传感器(组氨酸)激酶的基因(mesS)进行失活,产生了一个与mesR突变体一样无法在液体培养基中生长的突变体。DNA微阵列和实时逆转录PCR分析表明,先前显示与卡他莫拉菌在豚鼠鼻咽部持续存在能力有关的几个基因在mesR突变体中上调。在mesR突变体中上调的另外两个开放阅读框被证明编码小蛋白(LipA和LipB),它们与细菌粘附素具有氨基酸序列同源性,与细菌溶菌酶抑制剂具有结构同源性。lipA和lipB两者失活并不影响卡他莫拉菌O35E体外附着于人支气管上皮细胞系的能力。纯化的重组LipA和LipB融合蛋白在体外和唾液中均显示出抑制人溶菌酶活性的作用。在人脱铁乳铁蛋白存在的情况下,lipA lipB缺失突变体比野生型亲本菌株对人溶菌酶杀伤更敏感。这是关于卡他莫拉菌产生溶菌酶抑制剂的首次报道。