Identification and phenotypic characterization of the cell-division protein CdpA

被引:37
作者
Altermann, E
Buck, BL
Cano, R
Klaenhammer, TR
机构
[1] N Carolina State Univ, SE Dairy Foods Res Ctr, Dept Food Sci, Raleigh, NC 27695 USA
[2] Calif Polytech State Univ San Luis Obispo, Environm Biotechnol Inst, San Luis Obispo, CA 93407 USA
关键词
lactic acid bacteria; lactobacillus; cell adhesion; stress tolerance; functional inactivation; cell wall; cell separation;
D O I
10.1016/j.gene.2004.08.004
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Analysis of the automated computer annotation of the early draft phase genome of Lactobacillus acidophilus NCFM revealed the previously discovered S-layer gene slpA and an additional partial ORF with weak similarities to S-layer proteins. The entire gene was sequenced to reveal a 1799-bp gene coding for 599 amino acids with a calculated molecular mass of 64.8 kDa. No transcription or translation signals could be determined in close proximity to the 5'-region. However, a strong putative terminator with a free energy of -16.84 kcal/mol was identified directly downstream of the gene. A PSI-Blast analysis showed similarities to members of S-layer proteins, cell-wall associated proteinases and hexosyl-transferases. Calculation of an unrooted phylogenetic tree with other examples of S-layer proteins and proteinases placed the deduced protein separately from both groups. A derivative of L. acidophilus NCFM was constructed by targeted integration into the gene. SDS-PAGE analysis of non-covalently linked proteins of the cell wall of the mutant, compared to the wild type, revealed the loss of a cell-surface protein. Phenotypic analyses of the mutant revealed significant changes in cell morphology, altered responses to various environmental stresses, and lowered cell adhesion. Based on the in silico and functional analyses, we ascertained that this protein plays a role in cell-wall processing during the growth and cell-cell separation and designated the gene as cell-division protein, cdpA. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:189 / 197
页数:9
相关论文
共 29 条
[1]  
Altermann Eric, 2003, OMICS A Journal of Integrative Biology, V7, P161, DOI 10.1089/153623103322246557
[2]   Surface display of foreign epitopes on the Lactobacillus brevis S-layer [J].
Åvall-Jääskeläinen, S ;
Kylä-Nikkilä, K ;
Kahala, M ;
Miikkulainen-Lahti, T ;
Palva, A .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (12) :5943-5951
[3]   DETECTION AND ACTIVITY OF LACTACIN-B, A BACTERIOCIN PRODUCED BY LACTOBACILLUS-ACIDOPHILUS [J].
BAREFOOT, SF ;
KLAENHAMMER, TR .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1983, 45 (06) :1808-1815
[4]   Improved prediction of signal peptides: SignalP 3.0 [J].
Bendtsen, JD ;
Nielsen, H ;
von Heijne, G ;
Brunak, S .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 340 (04) :783-795
[5]   The presence of two S-layer-protein-encoding genes is conserved among species related to Lactobacillus acidophilus [J].
Boot, HJ ;
Kolen, CPAM ;
Pot, B ;
Kersters, K ;
Pouwels, PH .
MICROBIOLOGY-SGM, 1996, 142 :2375-2384
[6]   S-LAYER PROTEIN OF LACTOBACILLUS-ACIDOPHILUS ATCC-4356 - PURIFICATION, EXPRESSION IN ESCHERICHIA-COLI, AND NUCLEOTIDE-SEQUENCE OF THE CORRESPONDING GENE [J].
BOOT, HJ ;
KOLEN, CPAM ;
VANNOORT, JM ;
POUWELS, PH .
JOURNAL OF BACTERIOLOGY, 1993, 175 (19) :6089-6096
[7]   IDENTIFICATION, CLONING, AND NUCLEOTIDE-SEQUENCE OF A SILENT S-LAYER PROTEIN GENE OF LACTOBACILLUS-ACIDOPHILUS ATCC-4356 WHICH HAS EXTENSIVE SIMILARITY WITH THE S-LAYER PROTEIN GENE OF THIS SPECIES [J].
BOOT, HJ ;
KOLEN, CPAM ;
POUWELS, PH .
JOURNAL OF BACTERIOLOGY, 1995, 177 (24) :7222-7230
[8]   ADHESION OF HUMAN LACTOBACILLUS-ACIDOPHILUS STRAIN-LB TO HUMAN ENTEROCYTE-LIKE CACO-2 CELLS [J].
CHAUVIERE, G ;
COCONNIER, MH ;
KERNEIS, S ;
FOURNIAT, J ;
SERVIN, AL .
JOURNAL OF GENERAL MICROBIOLOGY, 1992, 138 :1689-1696
[9]   Anchorless adhesins and invasins of Gram-positive bacteria: a new class of virulence factors [J].
Chhatwal, GS .
TRENDS IN MICROBIOLOGY, 2002, 10 (05) :205-208
[10]   Expression of fibronectin-binding protein FbpA modulates adhesion in Streptococcus gordonii [J].
Christie, J ;
McNab, R ;
Jenkinson, HF .
MICROBIOLOGY-SGM, 2002, 148 :1615-1625