Analyses of ampC gene expression in Serratia marcescens reveal new regulatory properties

Steven D. Mahlen, Stacey S. Morrow, Baha Abdalhamid, Nancy D. Hanson

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Serratia marcescens encodes an inducible, chromosomal β-lactamase, ampC. Studies addressing the regulation of inducible ampC genes have focused primarily on Enterobacter cloacae and Citrobacter freundii. The purpose of this study was to clone and sequence the ampC, ampR and intergenic region of S. marcescens and examine both inducible and basal level ampC expression. Sequence analysis of the S. marcescens ampC gene identified an extended 5′ untranslated region (UTR) of 126 nucleotides, which formed a prominent stem-loop structure. Induction of ampC expression required AmpR, and the start of transcription was determined using primer extension analysis. In vivo half-life analysis revealed that the half-life of the S. marcescens ampC transcript was 7 min. Confirmation of the in vivo half-life and the role of the stem-loop structure in the 5′ UTR was demonstrated by comparing transcript half-life and luciferase expression between a wild-type (WT) and a 5′ UTR stem-loop deletion mutant. These data demonstrated that the stem-loop structure was involved in transcript stability. Taken together, these findings indicate that constitutive expression of S. marcescens ampC is regulated by both transcriptional initiation and post-transcriptional events.

Original languageEnglish
Pages (from-to)791-802
Number of pages12
JournalJournal of Antimicrobial Chemotherapy
Volume51
Issue number4
DOIs
StatePublished - Apr 1 2003

Fingerprint

Serratia marcescens
Half-Life
5' Untranslated Regions
Gene Expression
Citrobacter freundii
Enterobacter cloacae
Intergenic DNA
Luciferases
Genes
Sequence Analysis
Nucleotides
Clone Cells

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Microbiology

Cite this

Analyses of ampC gene expression in Serratia marcescens reveal new regulatory properties. / Mahlen, Steven D.; Morrow, Stacey S.; Abdalhamid, Baha; Hanson, Nancy D.

In: Journal of Antimicrobial Chemotherapy, Vol. 51, No. 4, 01.04.2003, p. 791-802.

Research output: Contribution to journalArticle

Mahlen, Steven D. ; Morrow, Stacey S. ; Abdalhamid, Baha ; Hanson, Nancy D. / Analyses of ampC gene expression in Serratia marcescens reveal new regulatory properties. In: Journal of Antimicrobial Chemotherapy. 2003 ; Vol. 51, No. 4. pp. 791-802.
@article{39ce29ca57ce4079b4b1f20a504d7aa9,
title = "Analyses of ampC gene expression in Serratia marcescens reveal new regulatory properties",
abstract = "Serratia marcescens encodes an inducible, chromosomal β-lactamase, ampC. Studies addressing the regulation of inducible ampC genes have focused primarily on Enterobacter cloacae and Citrobacter freundii. The purpose of this study was to clone and sequence the ampC, ampR and intergenic region of S. marcescens and examine both inducible and basal level ampC expression. Sequence analysis of the S. marcescens ampC gene identified an extended 5′ untranslated region (UTR) of 126 nucleotides, which formed a prominent stem-loop structure. Induction of ampC expression required AmpR, and the start of transcription was determined using primer extension analysis. In vivo half-life analysis revealed that the half-life of the S. marcescens ampC transcript was 7 min. Confirmation of the in vivo half-life and the role of the stem-loop structure in the 5′ UTR was demonstrated by comparing transcript half-life and luciferase expression between a wild-type (WT) and a 5′ UTR stem-loop deletion mutant. These data demonstrated that the stem-loop structure was involved in transcript stability. Taken together, these findings indicate that constitutive expression of S. marcescens ampC is regulated by both transcriptional initiation and post-transcriptional events.",
author = "Mahlen, {Steven D.} and Morrow, {Stacey S.} and Baha Abdalhamid and Hanson, {Nancy D.}",
year = "2003",
month = "4",
day = "1",
doi = "10.1093/jac/dkg133",
language = "English",
volume = "51",
pages = "791--802",
journal = "Journal of Antimicrobial Chemotherapy",
issn = "0305-7453",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Analyses of ampC gene expression in Serratia marcescens reveal new regulatory properties

AU - Mahlen, Steven D.

AU - Morrow, Stacey S.

AU - Abdalhamid, Baha

AU - Hanson, Nancy D.

PY - 2003/4/1

Y1 - 2003/4/1

N2 - Serratia marcescens encodes an inducible, chromosomal β-lactamase, ampC. Studies addressing the regulation of inducible ampC genes have focused primarily on Enterobacter cloacae and Citrobacter freundii. The purpose of this study was to clone and sequence the ampC, ampR and intergenic region of S. marcescens and examine both inducible and basal level ampC expression. Sequence analysis of the S. marcescens ampC gene identified an extended 5′ untranslated region (UTR) of 126 nucleotides, which formed a prominent stem-loop structure. Induction of ampC expression required AmpR, and the start of transcription was determined using primer extension analysis. In vivo half-life analysis revealed that the half-life of the S. marcescens ampC transcript was 7 min. Confirmation of the in vivo half-life and the role of the stem-loop structure in the 5′ UTR was demonstrated by comparing transcript half-life and luciferase expression between a wild-type (WT) and a 5′ UTR stem-loop deletion mutant. These data demonstrated that the stem-loop structure was involved in transcript stability. Taken together, these findings indicate that constitutive expression of S. marcescens ampC is regulated by both transcriptional initiation and post-transcriptional events.

AB - Serratia marcescens encodes an inducible, chromosomal β-lactamase, ampC. Studies addressing the regulation of inducible ampC genes have focused primarily on Enterobacter cloacae and Citrobacter freundii. The purpose of this study was to clone and sequence the ampC, ampR and intergenic region of S. marcescens and examine both inducible and basal level ampC expression. Sequence analysis of the S. marcescens ampC gene identified an extended 5′ untranslated region (UTR) of 126 nucleotides, which formed a prominent stem-loop structure. Induction of ampC expression required AmpR, and the start of transcription was determined using primer extension analysis. In vivo half-life analysis revealed that the half-life of the S. marcescens ampC transcript was 7 min. Confirmation of the in vivo half-life and the role of the stem-loop structure in the 5′ UTR was demonstrated by comparing transcript half-life and luciferase expression between a wild-type (WT) and a 5′ UTR stem-loop deletion mutant. These data demonstrated that the stem-loop structure was involved in transcript stability. Taken together, these findings indicate that constitutive expression of S. marcescens ampC is regulated by both transcriptional initiation and post-transcriptional events.

UR - http://www.scopus.com/inward/record.url?scp=0037392674&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037392674&partnerID=8YFLogxK

U2 - 10.1093/jac/dkg133

DO - 10.1093/jac/dkg133

M3 - Article

VL - 51

SP - 791

EP - 802

JO - Journal of Antimicrobial Chemotherapy

JF - Journal of Antimicrobial Chemotherapy

SN - 0305-7453

IS - 4

ER -