Importance of cis determinants and nitrogenase activity in regulated stability of the Klebsiella pneumoniae nitrogenase structural gene mRNA

Holly Simon, Mark M. Gosink, Gary P. Roberts

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The Klebsiella pneumoniae nitrogen fixation (nif) mRNAs are unusually stable, with half-lives of 20 to 30 min under conditions favorable to nitrogen fixation (limiting nitrogen, anaerobiosis, temperatures of 30°C). Addition of O2 or fixed nitrogen or temperature increases to 37°C or more result in the dramatic destabilization of the nif mRNAs, decreasing the half- lives by a factor of 3 to 5. A plasmid expression system, independent of nif transcriptional regulation, was used to define cis determinants required for the regulated stability of the 5.2-kb nifHDKTY mRNA and to test the model suggested by earlier work that NifA is required in trans to stabilize nif mRNA under nif-derepressing conditions. O2 regulation of nifHDKTY mRNA stability is impaired in a plasmid containing a deletion of a 499-bp region of nifH, indicating that a site(s) required for the O2-regulated stability of the mRNA is located within this region. The simple model suggested from earlier work that NifA is required for stabilizing nif mRNA under conditions favorable for nitrogen fixation was disproved, and in its place, a more complicated model involving the sensing of nitrogenase activity as a component of the system regulating mRNA stability is proposed. Analysis of nifY mutants and overexpression suggests a possible involvement of the protein in this sensing process.

Original languageEnglish (US)
Pages (from-to)3751-3760
Number of pages10
JournalJournal of Bacteriology
Volume181
Issue number12
StatePublished - Jun 1999
Externally publishedYes

Fingerprint

Nitrogenase
Nitrogen Fixation
Klebsiella pneumoniae
Messenger RNA
Genes
RNA Stability
Plasmids
Nitrogen
Anaerobiosis
Temperature

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Importance of cis determinants and nitrogenase activity in regulated stability of the Klebsiella pneumoniae nitrogenase structural gene mRNA. / Simon, Holly; Gosink, Mark M.; Roberts, Gary P.

In: Journal of Bacteriology, Vol. 181, No. 12, 06.1999, p. 3751-3760.

Research output: Contribution to journalArticle

@article{ff5e54b842e84a7d83bf99b03446ff75,
title = "Importance of cis determinants and nitrogenase activity in regulated stability of the Klebsiella pneumoniae nitrogenase structural gene mRNA",
abstract = "The Klebsiella pneumoniae nitrogen fixation (nif) mRNAs are unusually stable, with half-lives of 20 to 30 min under conditions favorable to nitrogen fixation (limiting nitrogen, anaerobiosis, temperatures of 30°C). Addition of O2 or fixed nitrogen or temperature increases to 37°C or more result in the dramatic destabilization of the nif mRNAs, decreasing the half- lives by a factor of 3 to 5. A plasmid expression system, independent of nif transcriptional regulation, was used to define cis determinants required for the regulated stability of the 5.2-kb nifHDKTY mRNA and to test the model suggested by earlier work that NifA is required in trans to stabilize nif mRNA under nif-derepressing conditions. O2 regulation of nifHDKTY mRNA stability is impaired in a plasmid containing a deletion of a 499-bp region of nifH, indicating that a site(s) required for the O2-regulated stability of the mRNA is located within this region. The simple model suggested from earlier work that NifA is required for stabilizing nif mRNA under conditions favorable for nitrogen fixation was disproved, and in its place, a more complicated model involving the sensing of nitrogenase activity as a component of the system regulating mRNA stability is proposed. Analysis of nifY mutants and overexpression suggests a possible involvement of the protein in this sensing process.",
author = "Holly Simon and Gosink, {Mark M.} and Roberts, {Gary P.}",
year = "1999",
month = "6",
language = "English (US)",
volume = "181",
pages = "3751--3760",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Importance of cis determinants and nitrogenase activity in regulated stability of the Klebsiella pneumoniae nitrogenase structural gene mRNA

AU - Simon, Holly

AU - Gosink, Mark M.

AU - Roberts, Gary P.

PY - 1999/6

Y1 - 1999/6

N2 - The Klebsiella pneumoniae nitrogen fixation (nif) mRNAs are unusually stable, with half-lives of 20 to 30 min under conditions favorable to nitrogen fixation (limiting nitrogen, anaerobiosis, temperatures of 30°C). Addition of O2 or fixed nitrogen or temperature increases to 37°C or more result in the dramatic destabilization of the nif mRNAs, decreasing the half- lives by a factor of 3 to 5. A plasmid expression system, independent of nif transcriptional regulation, was used to define cis determinants required for the regulated stability of the 5.2-kb nifHDKTY mRNA and to test the model suggested by earlier work that NifA is required in trans to stabilize nif mRNA under nif-derepressing conditions. O2 regulation of nifHDKTY mRNA stability is impaired in a plasmid containing a deletion of a 499-bp region of nifH, indicating that a site(s) required for the O2-regulated stability of the mRNA is located within this region. The simple model suggested from earlier work that NifA is required for stabilizing nif mRNA under conditions favorable for nitrogen fixation was disproved, and in its place, a more complicated model involving the sensing of nitrogenase activity as a component of the system regulating mRNA stability is proposed. Analysis of nifY mutants and overexpression suggests a possible involvement of the protein in this sensing process.

AB - The Klebsiella pneumoniae nitrogen fixation (nif) mRNAs are unusually stable, with half-lives of 20 to 30 min under conditions favorable to nitrogen fixation (limiting nitrogen, anaerobiosis, temperatures of 30°C). Addition of O2 or fixed nitrogen or temperature increases to 37°C or more result in the dramatic destabilization of the nif mRNAs, decreasing the half- lives by a factor of 3 to 5. A plasmid expression system, independent of nif transcriptional regulation, was used to define cis determinants required for the regulated stability of the 5.2-kb nifHDKTY mRNA and to test the model suggested by earlier work that NifA is required in trans to stabilize nif mRNA under nif-derepressing conditions. O2 regulation of nifHDKTY mRNA stability is impaired in a plasmid containing a deletion of a 499-bp region of nifH, indicating that a site(s) required for the O2-regulated stability of the mRNA is located within this region. The simple model suggested from earlier work that NifA is required for stabilizing nif mRNA under conditions favorable for nitrogen fixation was disproved, and in its place, a more complicated model involving the sensing of nitrogenase activity as a component of the system regulating mRNA stability is proposed. Analysis of nifY mutants and overexpression suggests a possible involvement of the protein in this sensing process.

UR - http://www.scopus.com/inward/record.url?scp=0032983098&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032983098&partnerID=8YFLogxK

M3 - Article

C2 - 10368150

AN - SCOPUS:0032983098

VL - 181

SP - 3751

EP - 3760

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 12

ER -