The psychrotrophic bacterium Yersinia enterocolitica requires expression of pnp, the gene for polynucleotide phosphorylase, for growth at low temperature (5 degrees C).
The psychrotrophic bacterium Yersinia enterocolitica is characterized by temperature-dependent adaptations. To investigate Y. enterocolitica genes involved in cold adaptation, a mutant restricted in its ability to grow at 5 degrees C was isolated from a transposon mutant library. The transposon insertion site in this psychrotrophy-defective (PD) mutant mapped 16 bp upstream of an open reading frame whose predicted amino acid sequence showed 93% similarity with the Escherichia coli exoribonuclease polynucleotide phosphorylase (PNPase), encoded by pnp. Expression of this gene was blocked in the PD mutant. However, the introduction of a second copy of pnp, including 0.33 kbp sequences upstream of its coding region, into the chromosome of the PD mutant restored pnp expression as well as the ability to grow at 5 degrees C. Furthermore, the expression of pnp appeared to be temperature dependent: in the parental Y. enterocolitica strain, the levels of both pnp mRNA and PNPase were 1.6-fold higher at 5 degrees C compared with 30 degrees C. A similarly enhanced level of PNPase at 5 degrees C was observed in the merodiploid recombinant strain, which indicates that the 0.33 kbp region upstream of pnp harboured a cold-inducible promoter. A putative cold shock promoter motif (ATTGG) was observed in this region.
Goverde
R L
RL
University of Utrecht, Faculty of Veterinary Medicine, Department of the Science of Food of Animal Origin, The Netherlands. goverde@vvdo.dgk.ruu.nl
Huis in't Veld
J H
JH
Kusters
J G
JG
Mooi
F R
FR
D001483:Base Sequence; D003080:Cold Temperature; D004251:DNA Transposable Elements; D018628:Gene Dosage; D015870:Gene Expression; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011117:Polyribonucleotide Nucleotidyltransferase / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D011995:Recombination, Genetic; D015008:Yersinia enterocolitica / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification
D004251:DNA Transposable Elements; D012329:RNA, Bacterial; D012333:RNA, Messenger; D011117:Polyribonucleotide Nucleotidyltransferase