Liu Dali, Yumoto Hiromichi, Murakami Keiji, Hirota Katsuhiko, Ono Tsuneko, Nagamune Hideaki, Kayama Shizuo, Matsuo Takashi, Miyake Yoichiro
Department of Microbiology, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kumamoto-cho, Tokushima 770-8504, Japan.
Mol Microbiol. 2008 Jun;68(5):1268-82. doi: 10.1111/j.1365-2958.2008.06232.x. Epub 2008 Apr 11.
Streptococcus intermedius histone-like DNA-binding protein (Si-HLP) is a homodimeric protein and, conserved with Escherichia coli HU, a well-documented nucleoid-associated protein (NAP). In E. coli, HU plays important roles as both structural and regulatory factors, but it is not essential for E. coli viability. Streptococcal HLP has been found to bind host cells and induce cytokine production, but its physiological role remains poorly defined. In the present study, using gene insertion knockout and tetracycline-regulated antisense RNA expression techniques, we determined whether Si-HLP is essential for bacterial viability and normal growth in S. intermedius. The Si-HLP-downregulated S. intermedius strain showed alterations in its morphology and surface properties. Downregulation of Si-HLP led to an expanded nucleoid to fill the intracellular space. Transcription levels of several genes, including virulence-associated factors, were found to be activated or repressed in the antisense Si-hlp RNA-expressing strain by real-time PCR and reverse-transcription PCR. Collectively, these data suggest that Si-HLP serves as an essential NAP governing the nucleoid architecture and controlling the gene transcription profile in S. intermedius.
中间型链球菌组蛋白样DNA结合蛋白(Si-HLP)是一种同二聚体蛋白,与大肠杆菌HU(一种有充分文献记载的类核相关蛋白(NAP))保守。在大肠杆菌中,HU作为结构和调节因子发挥重要作用,但对大肠杆菌的生存力并非必不可少。已发现链球菌HLP可结合宿主细胞并诱导细胞因子产生,但其生理作用仍不清楚。在本研究中,我们使用基因插入敲除和四环素调控的反义RNA表达技术,确定Si-HLP对中间型链球菌的细菌生存力和正常生长是否必不可少。Si-HLP下调的中间型链球菌菌株在形态和表面特性上出现了改变。Si-HLP的下调导致类核扩大以填充细胞内空间。通过实时PCR和逆转录PCR发现,在表达反义Si-hlp RNA的菌株中,包括毒力相关因子在内的几个基因的转录水平被激活或抑制。总体而言,这些数据表明Si-HLP作为一种必需的NAP,在中间型链球菌中调控类核结构并控制基因转录谱。