Identification and characterization of a peptidoglycan hydrolase, MurA, of Listeria monocytogenes, a muramidase needed for cell separation.
A novel cell wall hydrolase encoded by the murA gene of Listeria monocytogenes is reported here. Mature MurA is a 66-kDa cell surface protein that is recognized by the well-characterized L. monocytogenes-specific monoclonal antibody EM-7G1. MurA displays two characteristic features: (i) an N-terminal domain with homology to muramidases from several gram-positive bacterial species and (ii) four copies of a cell wall-anchoring LysM repeat motif present within its C-terminal domain. Purified recombinant MurA produced in Escherichia coli was confirmed to be an authentic cell wall hydrolase with lytic properties toward cell wall preparations of Micrococcus lysodeikticus. An isogenic mutant with a deletion of murA that lacked the 66-kDa cell wall hydrolase grew as long chains during exponential growth. Complementation of the mutant strain by chromosomal reintegration of the wild-type gene restored expression of this murein hydrolase activity and cell separation levels to those of the wild-type strain. Studies reported herein suggest that the MurA protein is involved in generalized autolysis of L. monocytogenes.
Carroll
Shannon A
SA
Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland 20742, USA.
Hain
Torsten
T
Technow
Ulrike
U
Darji
Ayub
A
Pashalidis
Philippos
P
Joseph
Sam W
SW
Chakraborty
Trinad
T
D000595:Amino Acid Sequence; D002473:Cell Wall / Q000201:enzymology; D003001:Cloning, Molecular; D016384:Consensus Sequence; D017353:Gene Deletion; D016680:Genome, Bacterial; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000472:pathogenicity; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009113:Muramidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D020079:Terminal Repeat Sequences
D011994:Recombinant Proteins; D009113:Muramidase; D009238:N-Acetylmuramoyl-L-alanine Amidase