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The characterization of DNA-binding protein from Streptococcus intermediusand elucidate its virulence mechanism

中间链球菌 DNA 结合蛋白的表征并阐明其毒力机制

基本信息

批准号：
14571788
负责人：
HIROTA Katsuhiko
金额：
$ 1.92万
依托单位：
Tokushima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

The histone-like proteins (HLPs) play a role in DNA replication, recombination, repair, Mutransposition, and regulation of gene transcription. In addition to their roles in cellular processes, HLPs are considered as an adhesin of some bacteria such as Streptococcus pyogenes. In the present study we cloned a gene encoding a histone-l ike protein (HLPSi) from Streptococcus intermediusUNS46. The deduced amino acid sequence of HLPSi showed 90.1%, 71.6%, 62.5%, 60.7%, and 36.4% identity with a histone-like protein of S. pyogenes A374, Bacillus subtilis, Escherichis coli HU-1, E.coli HU-2, and Mycobacterium leprae TN, respectively. HLPSi was rich in alanine (20.9%), lysine (13.2%), and arginine (4.4%) and devoid of tryptophan, tyrosine, histidine, and cysteine. The recombinant protein was expressed in E.coli JM109 by use of the pGEX expression system. Comparison of the deduced amino acid sequence of HLPSi with known protein sequences deposited in databases revealed that this protein had 90.1%, 71.6%, 62.5%, 60.7% and 36.4% identity over 91 aa residues with histone-like DNA-binding proteins of S.pyogenes A374, Bacillus subtilis, E.coli HU-1 and HU-2, and M. leprae TN, respectively. Anti HLPSi antibody reacted the whole cells of S. intermedius NCDO2227, S.pyogenesA374, S anginosus TW1751. S.constellatus NCD02226, S.inutans lngbritt, S.sobrinus 0MZ65, S.sanguinis ATCC10556, S. sanguinis ST6, S.gordonii Challis, S.salivarius HT9R, S. salivarius NCTC8168, S.mitis ATCC15913, S. aureus Smith, S. aureus SMF-4. Although the export mechanism of HLPSI is still unknown, this is perhaps due to the release of the protein during induced alterations in membrane permeability, leading to the release of this protein along with other cytoplasmic proteins. Further experiments are necessary to define the various aspects of the binding function of HLPSi to host cells.

组蛋白样蛋白 (HLP) 在 DNA 复制、重组、修复、Mu转座和基因转录调节中发挥作用。除了在细胞过程中的作用外，HLP 还被认为是某些细菌（例如化脓性链球菌）的粘附素。在本研究中，我们从中间链球菌UNS46中克隆了编码组蛋白样蛋白（HLPSi）的基因。 HLPSi 的推导氨基酸序列与化脓性链球菌 A374、枯草芽孢杆菌、大肠杆菌 HU-1、大肠杆菌 HU-2 和麻风分枝杆菌 TN 的组蛋白样蛋白分别具有 90.1%、71.6%、62.5%、60.7% 和 36.4% 的一致性。 HLPSi 富含丙氨酸（20.9%）、赖氨酸（13.2%）和精氨酸（4.4%），不含色氨酸、酪氨酸、组氨酸和半胱氨酸。利用pGEX表达系统在大肠杆菌JM109中表达重组蛋白。将HLPSi推导的氨基酸序列与数据库中已知的蛋白质序列进行比较表明，该蛋白质在91个氨基酸残基上与化脓性链球菌A374、枯草芽孢杆菌、大肠杆菌HU-1和HU-2以及麻风杆菌的组蛋白样DNA结合蛋白具有90.1％、71.6％、62.5％、60.7％和36.4％的同一性分别为TN。抗HLPSi抗体与中间链球菌NCDO2227、化脓链球菌A374、咽峡链球菌TW1751的全细胞反应。 S.constellatus NCD02226、S.inutans lngbritt、S.sobrinus 0MZ65、S.sanguinis ATCC10556、S.sanguinis ST6、S.gordonii Challis、S.salivarius HT9R、S.salivarius NCTC8168、S.mitis ATCC15913、S.aureus Smith、S.金黄色葡萄球菌SMF-4。尽管 HLPSI 的输出机制仍不清楚，但这可能是由于在膜通透性诱导改变过程中蛋白质的释放，导致该蛋白质与其他细胞质蛋白质一起释放。需要进一步的实验来确定 HLPSi 与宿主细胞结合功能的各个方面。