Cloning and sequencing of two genes, prtA and prtB, from Myxococcus xanthus, encoding PrtA and PrtB proteases, both of which are required for the protease activity.
The sequence of a 1955-bp TaqI DNA fragment from Myxococcus xanthus was determined. This fragment contains two complete genes, designated prtA and prtB. The prtA and prtB ORFs extend over 828 and 798 bp, respectively. They are separated only by 3 nt and appear to be present in a polycistronic transcriptional unit. A typical lipoprotein signal sequence is present at the N terminus of the two deduced polypeptides. The aa sequence of PrtA shows a high degree of identity to the region adjacent to the Ser residue belonging to the catalytic triad of serine proteases from Staphylococcus aureus and Enterococcus faecalis. It also exhibits features characteristic of trypsin-like serine proteases in that it contains the same pattern of variable and conserved regions. The deduced aa sequence of PrtB reveals a signature zinc-binding consensus motif (HEXXHXXGXXH/Met-turn) characteristic of the class of metalloproteases called metzincins. Plasmids containing prtA, prtB, or both were constructed. Protease activity studies of Escherichia coli clones containing these plasmids showed that both genes are necessary for this activity, whatever their cis or trans position. As prtB produces a putative membrane-bound lipoprotein of 266 aa, the protease activation must occur at the membrane level.
Quillet
L
L
Laboratoire de Microbiologie du froid, I.F.R. 61 CNRS INSERM, Faculté des Sciences, Université de Rouen, Mont Saint-Aignan, France. quillet@coloris.univ-rouen.fr
Bensmail
L
L
Barray
S
S
Guespin-Michel
J
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics*; D005798:Genes, Bacterial; D008074:Lipoproteins / Q000235:genetics; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016941:Myxococcus xanthus / Q000201:enzymology* / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics*; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; D008074:Lipoproteins; D010450:Endopeptidases; D012697:Serine Endopeptidases; D008666:Metalloendopeptidases; C079508:PrtA metalloprotease; C099750:PrtB protein, bacteria