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PMID: 1622271 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular analyses of the lactococcin A gene cluster from Lactococcus lactis subsp. lactis biovar diacetylactis WM4.

Applied and environmental microbiology ·Vol. 58 ·No. 6 ·1992-06-00 ·Pages 1952-61

Stoddard GW, Petzel JP, van Belkum MJ, Kok J, McKay LL

Abstract

The genes responsible for bacteriocin production and immunity in Lactococcus lactis subsp. lactis biovar diacetylactis WM4 were localized and characterized by DNA restriction fragment deletion, subcloning, and nucleotide sequence analysis. The nucleotide sequence of a 5.6-kb AvaII restriction fragment revealed a cluster with five complete open reading frames (ORFs) in the same orientation. DNA and protein homology analyses, combined with deletion and Tn5 insertion mutagenesis, implicated four of the ORFs in the production of and immunity to lactococcin A. The last two ORFs in the cluster were the lactococcin A structural and immunity genes, lcnA and lciA. The two ORFs immediately upstream of lcnA and lciA were designated lcnC and lcnD, and the proteins that they encoded showed similarities to proteins of signal sequence-independent secretion systems. lcnC encodes a protein of 716 amino acids that could belong to the HlyB family of ATP-dependent membrane translocators. LcnC contains an ATP binding domain in a conserved C-terminal stretch of approximately 200 amino acids and three putative hydrophobic segments in the N terminus. The lcnD product, LcnD, of 474 amino acids, is essential for lactococcin A expression and shows structural similarities to HlyD and its homologs. On the basis of these results, a secretion apparatus that is essential for the full expression of active lactococcin A is postulated.

MeSH Terms
Adenosine Triphosphate/metabolism Amino Acid Sequence Bacterial Proteins/genetics Bacteriocins/biosynthesis Base Sequence Carrier Proteins/genetics DNA, Bacterial/genetics Genes, Bacterial Lactococcus lactis/genetics,immunology,metabolism Membrane Proteins/genetics Molecular Sequence Data Multigene Family Open Reading Frames Plasmids Restriction Mapping Sequence Homology, Nucleic Acid
Chemicals
Bacterial Proteins Bacteriocins Carrier Proteins DNA, Bacterial LcnD protein, Lactococcus lactis Membrane Proteins lactococcin A Adenosine Triphosphate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Stoddard G W
Department of Food Science and Nutrition, University of Minnesota, St. Paul 55108.
Petzel J P
van Belkum M J
Kok J
McKay L L
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.58.6.1952-1961.1992
Published
1992-06-00
Pages
1952-61
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC195709
Subset
IM
Databases
GENBANK
M55228, M55229, M55230, M55231, M55232, M55233, M55234, M90969, S39189, S39231
External Links
PubMed source DOI: 10.1128/aem.58.6.1952-1961.1992 PMC Full Text View journal details
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