Controlled expression of CluA in Lactococcus lactis and its role in conjugation.
CluA is a 136 kDa surface-bound protein encoded by the chromosomally located sex factor of Lactococcus lactis MG1363 and is associated with cell aggregation linked to high-frequency transfer of the sex factor. To further investigate the involvement of CluA in these phenomena, the cluA gene was cloned on a plasmid, downstream from the lactococcal nisA promoter. In a sex-factor-negative MG1363 derivative, nisin-controlled CluA expression resulted in aggregation, despite the absence of the other genes of the sex factor. Therefore, CluA is the only sex factor component responsible for aggregation. The direct involvement of CluA in the establishment of cell-to-cell contact for aggregate formation was observed by electron microscopy using immunogold-labelled CluA antibodies. Inactivation of cluA in an MG1363 background led to a dramatic decrease in sex factor conjugation frequency compared to the parental strain. Increasing levels of CluA expressed in trans in the cluA-inactivated donor strain facilitated a gradual restoration of conjugation frequency, reaching that of the parental strain. In conclusion, CluA is essential for efficient sex factor transfer in conjugation of L. lactis.
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Régis
R
Stentz
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Karen
K
Jury
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Tracy
T
Eaton
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Mary
M
Parker
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Arjan
A
Narbad
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Mike
M
Gasson
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.
Claire
C
Shearman
D000818:Animals; D000907:Antibodies, Bacterial; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics; D005144:F Factor / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D007150:Immunohistochemistry; D007763:Lac Operon; D013294:Lactococcus lactis / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D008854:Microscopy, Electron; D009561:Nisin / Q000235:genetics; D011401:Promoter Regions, Genetic
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009561:Nisin