Cloning and nucleotide sequence of the csp1 gene encoding PS1, one of the two major secreted proteins of Corynebacterium glutamicum: the deduced N-terminal region of PS1 is similar to the Mycobacterium antigen 85 complex.
Two proteins, PS1 and PS2, were detected in the culture medium of Corynebacterium glutamicum and are the major proteins secreted by this bacterium. No enzymatic activity was identified for either of the two proteins. Immunologically cross-reacting proteins were found in a variety of C. glutamicum strains but not in the coryneform Arthrobacter aureus. The gene encoding PS1, csp1, was cloned in lambda gt11 using polyclonal antibodies raised against PS1 to screen for producing clones. The csp1 gene was expressed in Escherichia coli, presumably from its own promoter, and directed the synthesis of two proteins recognized by anti-PS1 antibodies. The major protein band, of lower M(r), was detected in the periplasmic fraction. It had the same M(r) as the PS1 protein band detected in the supernatant of C. glutamicum cultures and presumably corresponds to the mature form of PS1. The minor protein band appears to be the precursor form of PS1. The nucleotide sequence of the csp1 gene was determined and contained an open reading frame encoding a polypeptide with a calculated molecular weight of 70,874, with a putative signal peptide with a molecular weight of 4411. This is consistent with the M(r) determined for PS1 from C. glutamicum culture supernatant and E. coli whole-cell extracts. The NH2-half of the deduced amino acid is similar (about 33% identical residues and 52% including similar residues) to the secreted antigen 85 protein complex of Mycobacterium. The csp1 gene in C. glutamicum was disrupted without any apparent effect on growth or viability.
Joliff
G
G
Centre Orsan de Recherche en Biotechnologie, Courtaboeuf, France.
Mathieu
L
L
Hahn
V
V
Bayan
N
N
Duchiron
F
F
Renaud
M
M
Schechter
E
E
Leblon
G
G
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D010582:Bacteriophage lambda; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000235:genetics*; D004269:DNA, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D015698:Genomic Library; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics; D015183:Restriction Mapping; D017385:Sequence Homology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D021382:Protein Sorting Signals; C077000:PS1 protein, Corynebacterium glutamicum