Transcriptional analysis of long-term adaptation of Yersinia enterocolitica to low-temperature growth.
To analyze the transcriptional response of Yersinia enterocolitica cells to prolonged growth at low temperature, a collection of luxCDABE transposon mutants was cultivated in parallel at optimal (30 degrees C) and suboptimal (10 degrees C) temperatures and screened for enhanced promoter activities during growth until entering stationary phase. Among 5,700 Y. enterocolitica mutants, 42 transcriptional units were identified with strongly enhanced or reduced promoter activity at 10 degrees C compared to 30 degrees C, and changes in their transcriptional levels over time were measured. Green fluorescent protein fusions to 10 promoter regions confirmed the data. The temporal order of induction of the temperature-responsive genes of Y. enterocolitica was deduced, starting with the expression of cold shock genes cspA and cspB and the elevated transcription of a glutamate-aspartate symporter. Subsequently, cold-adapted cells drastically up-regulated genes encoding environmental sensors and regulators, such as UhpABC, ArcA, and methyl-accepting chemotaxis protein I (MCPI). Among the most prominent cold-responsive elements that were transcriptionally induced during growth in early and middle exponential phase are the insecticidal toxin genes tcaA and tcaB, as well as genes involved in flagellar synthesis and chemotaxis. The expression pattern of the late-exponential- to early-stationary-growth phase is dominated by factors involved in biodegradative metabolism, namely, a histidine ammonia lyase, three enzymes responsible for uptake and utilization of glycogen, the urease complex, and a subtilisin-like protease. Double-knockout mutants and complementation studies demonstrate inhibitory effects of MCPI and UhpC on the expression of a putative hemolysin transporter. The data partially delineate the spectrum of gene expression of Y. enterocolitica at environmental temperatures, providing evidence that an as-yet-unknown insect phase is part of the life cycle of this human pathogen.
Bresolin
Geraldine
G
Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Abteilung Mikrobiologie, Technische Universität München, Weihenstephaner Berg 3, D-85354 Freising, Germany.
Neuhaus
Klaus
K
Scherer
Siegfried
S
Fuchs
Thilo M
TM
D000222:Adaptation, Physiological; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D002633:Chemotaxis / Q000235:genetics; D003080:Cold Temperature; D004251:DNA Transposable Elements; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005809:Genes, Regulator; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D027981:Symporters / Q000235:genetics; D013997:Time Factors; D015008:Yersinia enterocolitica / Q000235:genetics / Q000254:growth & development*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004251:DNA Transposable Elements; D027981:Symporters; C062260:cold shock protein CS7.4, Bacteria; C077188:cold-shock protein CspB, Bacteria; D049452:Green Fluorescent Proteins; D008156:Luciferases