Structure, function, and fate of the BlaR signal transducer involved in induction of beta-lactamase in Bacillus licheniformis

The membrane-spanning protein BlaR is essential for the induction of beta-lactamase in Bacillus licheniformis. Its nature and location were confirmed by the use of an antiserum specific for its carboxy-terminal penicillin sensor, its function was studied by genetic dissection, and the structure of the penicillin sensor was derived from hydrophobic cluster analysis of the amino acid sequence by using, as a reference, the class A beta-lactamases with known three-dimensional structures. During the first 2 h after the addition of the beta-lactam inducer, full-size BlaR, bound to the plasma membrane, is produced, and then beta-lactamase is produced. By 2 h after induction, BlaR is present in various (membrane-bound and cytosolic) forms, and there is a gradual decrease in beta-lactamase production. The penicillin sensors of BlaR and the class D beta-lactamases show strong similarities in primary structures. They appear to have the same basic spatial disposition of secondary structures as that of the class A beta-lactamases, except that they lack several alpha helices and, therefore, have a partially uncovered five-stranded beta sheet and a more readily accessible active site. Alterations of BlaR affecting conserved secondary structures of the penicillin sensor and specific sites of the transducer annihilate beta-lactamase inducibility.

[1]  J M Ghuysen,et al.  Comparison of the sequences of class A beta-lactamases and of the secondary structure elements of penicillin-recognizing proteins , 1991, Antimicrobial Agents and Chemotherapy.

[2]  J. Frère,et al.  Mechanism of acyl transfer by the class A serine beta-lactamase of Streptomyces albus G. , 1991, The Biochemical journal.

[3]  O. Herzberg,et al.  Refined crystal structure of beta-lactamase from Staphylococcus aureus PC1 at 2.0 A resolution. , 1991, Journal of molecular biology.

[4]  J. Ghuysen,et al.  Identification of BlaR, the signal transducer for beta-lactamase production in Bacillus licheniformis, as a penicillin-binding protein with strong homology to the OXA-2 beta-lactamase (class D) of Salmonella typhimurium , 1990, Journal of bacteriology.

[5]  J M Ghuysen,et al.  The active-site-serine penicillin-recognizing enzymes as members of the Streptomyces R61 DD-peptidase family. , 1988, The Biochemical journal.

[6]  J. Lampen,et al.  Differential transcription of the bla regulatory region during induction of β‐lactamase in Bacillus licheniformis , 1988, FEBS letters.

[7]  J. Mornon,et al.  Hydrophobic cluster analysis: An efficient new way to compare and analyse amino acid sequences , 1987, FEBS letters.

[8]  T. Kobayashi,et al.  A second regulatory gene, blaR1, encoding a potential penicillin-binding protein required for induction of beta-lactamase in Bacillus licheniformis , 1987, Journal of bacteriology.

[9]  J M Ghuysen,et al.  The crystal structure of the beta-lactamase of Streptomyces albus G at 0.3 nm resolution. , 1987, The Biochemical journal.

[10]  J. Lampen,et al.  Transcriptional analysis of beta-lactamase regulation in Bacillus licheniformis , 1986, Journal of bacteriology.

[11]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[12]  C. Richardson,et al.  A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. W. Davis,et al.  Efficient isolation of genes by using antibody probes. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Beckwith,et al.  Regulation of a membrane component required for protein secretion in escherichia coli , 1982, Cell.

[15]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Sherratt,et al.  Analysis by transformation of the penicillinase system in Bacillus licheniformis. , 1973, Journal of general microbiology.

[17]  M. Sargent Rapid fixed-time assay for penicillinase , 1968, Journal of bacteriology.

[18]  D. Dubnau,et al.  The genetics of Bacillus licheniformis penicillinase: a preliminary analysis from studies on mutation and inter-strain and intra-strain transformations. , 1965, Journal of general microbiology.

[19]  J. Ghuysen,et al.  Serine beta-lactamases and penicillin-binding proteins. , 1991, Annual review of microbiology.

[20]  Bernard Henrissat,et al.  Structural homology among the peroxidase enzyme family revealed by hydrophobic cluster analysis , 1990, Proteins.

[21]  J. W. Dale,et al.  The dimeric nature of an R-factor mediated beta-lactamase. , 1976, Biochemical and biophysical research communications.