The cold shock response of the psychrotrophic bacterium Pseudomonas fragi involves four low-molecular-mass nucleic acid-binding proteins.
The psychrotrophic bacterium Pseudomonas fragi was subjected to cold shocks from 30 or 20 to 5 degrees C. The downshifts were followed by a lag phase before growth resumed at a characteristic 5 degrees C growth rate. The analysis of protein patterns by two-dimentional gel electrophoresis revealed overexpression of 25 or 17 proteins and underexpression of 12 proteins following the 30- or 20-to-5 degrees C shift, respectively. The two downshifts shared similar variations of synthesis of 20 proteins. The kinetic analysis distinguished the induced proteins into cold shock proteins (Csps), which were rapidly but transiently overexpressed, and cold acclimation proteins (Caps), which were more or less rapidly induced but still overexpressed several hours after the downshifts. Among the cold-induced proteins, four low-molecular-mass proteins, two of them previously characterized as Caps (CapA and CapB), and heat acclimation proteins (Haps) as well as heat shock proteins (Hsps) for the two others (TapA and TapB) displayed higher levels of induction. Partial amino acid sequences, obtained by microsequencing, were used to design primers to amplify by PCR the four genes and then determine their nucleotide sequences. A BamHI-EcoRI restriction fragment of 1.9 kb, containing the complete coding sequence for capB, was cloned and sequenced. The four peptides belong to the family of small nucleic acid-binding proteins as CspA, the major Escherichia coli Csp. They are likely to play a major role in the adaptative response of P. fragi to environmental temperature changes.
Michel
V
V
Station de Recherches sur la Viande, Unité de Recherches de Microbiologie, Institut National de la Recherche Agronomique de Theix, Saint-Genès-Champanelle, France.
Lehoux
I
I
Depret
G
G
Anglade
P
P
Labadie
J
J
Hebraud
M
M
D000220:Adaptation, Biological / Q000235:genetics*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D003001:Cloning, Molecular; D003080:Cold Temperature; D004268:DNA-Binding Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009696:Nucleic Acids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011549:Pseudomonas / Q000235:genetics*; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; C108302:CapA protein, bacteria; C109404:CapB protein, Pseudomonas fragi; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D009696:Nucleic Acids; C109405:TapA protein, Pseudomonas; C109406:TapB protein, Pseudomonas fragi