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PMID: 8141562 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Sequence analysis of PER-1 extended-spectrum beta-lactamase from Pseudomonas aeruginosa and comparison with class A beta-lactamases.

Antimicrobial agents and chemotherapy ·Vol. 38 ·No. 1 ·1994-01-00 ·Pages 104-14

Nordmann P, Naas T

Abstract

We have determined the nucleotide sequence (EMBL accession number, Z 21957) of the cloned chromosomal PER-1 extended-spectrum beta-lactamase gene from a Pseudomonas aeruginosa RNL-1 clinical isolate, blaPER-1 corresponds to a 924-bp open reading frame which encodes a polypeptide of 308 amino acids. This open reading frame is preceded by a -10 and a -35 region consistent with a putative P. aeruginosa promoter. Primer extension analysis of the PER-1 mRNA start revealed that this promoter was active in P. aeruginosa but not in Escherichia coli, in which PER-1 expression was driven by vector promoter sequences. N-terminal sequencing identified the PER-1 26-amino-acid leader peptide and enabled us to calculate the molecular mass (30.8 kDa) of the PER-1 mature form. Analysis of the percent GC content of blaPER-1 and of its 5' upstream sequences, as well as the codon usage for blaPER-1, indicated that blaPER-1 may have been inserted into P. aeruginosa genomic DNA from a nonpseudomonad bacterium. The PER-1 gene showed very low homology with other beta-lactamase genes at the DNA level. By using computer methods, assessment of the extent of identity between PER-1 and 10 beta-lactamase amino acid sequences indicated that PER-1 is a class A beta-lactamase. PER-1 shares around 27% amino acid identity with the sequenced extended-spectrum beta-lactamases of the TEM-SHV series and MEN-1 from Enterobacteriaceae species. The use of parsimony methods showed that PER-1 is not more closely related to gram-negative than to gram-positive bacterial class A beta-lactamases. Surprisingly, among class A beta-lactamases, PER-1 was most closely related to the recently reported CFXA from Bacteroides vulgatus, with which it shared 40% amino acid identity. This work indicates that non-Enterobacteriaceae species such as P. aeruginosa may possess class A extended-spectrum beta-lactamase genes possibly resulting from intergeneric DNA transfer.

Related Genes
bla PER-1
MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Culture Media DNA, Bacterial/analysis,isolation & purification Escherichia coli/genetics Molecular Sequence Data Molecular Weight Open Reading Frames Plasmids Pseudomonas aeruginosa/enzymology,genetics Sequence Homology, Nucleic Acid beta-Lactamases/analysis,genetics
Chemicals
Culture Media DNA, Bacterial beta-Lactamases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Nordmann P
Service de Microbiologie, Hôpital Raymond Poincaré, Faculté de Médecine Paris-Ouest, Université Paris, Garches, France.
Naas T
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Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
0066-4804
DOI
10.1128/AAC.38.1.104
Published
1994-01-00
Pages
104-14
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC284404
Subset
IM
Databases
GENBANK
Z21957
External Links
PubMed source DOI: 10.1128/AAC.38.1.104 PMC Full Text View journal details
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