Internal promoter characterization and expression of the Deinococcus radiodurans pprI-folP gene cluster

Guanjun Gao, Donghai Le, Lifen Huang, Huiming Lu, Issay Narumi, Yuejin Hua

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

PprI is a general gene switch responsible for the extraordinary radioresistance of Deinococcus radiodurans. From NCBI DNA sequence analysis, it was predicted that the translation start codon of the downstream folP (DR0168) gene overlaps the pprI (DR0167) stop codon, suggesting that these genes may form an operon. In this study, we show that a mutant containing an inserted sequence in folP does not grow unless folate is added to the medium, but is not affected in extreme radioresistance, whereas a pprI disruptant strain could grow in the absence of folate. It was found that expression of a pprI-lacZ fusion is constitutive and unaltered following ionizing radiation as is the production of the PprI protein. PprI protein is not expressed if its promoter is deleted and the transcription from the entire pprI promoter is essential for radioresistance of D. radiodurans. However, the deletion of pprI promoter has no effect on the expression of the folP-lacZ fusion. Primer extension analysis of the folP promoter region shows that folP is transcribed from its own promoter located within the pprI structural gene. All these results do support neither the existence of a pprI-folP operon nor a regulatory role of FolP in pprI expression.

Original languageEnglish (US)
Pages (from-to)195-201
Number of pages7
JournalFEMS Microbiology Letters
Volume257
Issue number2
DOIs
StatePublished - Apr 1 2006

Fingerprint

Deinococcus
Multigene Family
Operon
Folic Acid
Switch Genes
Genes
Initiator Codon
Terminator Codon
Ionizing Radiation
DNA Sequence Analysis
Genetic Promoter Regions
Proteins

Keywords

  • Deincoccus radiodurans
  • folP
  • lacZ-fusion
  • pprI
  • Promoter
  • Western blot

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Internal promoter characterization and expression of the Deinococcus radiodurans pprI-folP gene cluster. / Gao, Guanjun; Le, Donghai; Huang, Lifen; Lu, Huiming; Narumi, Issay; Hua, Yuejin.

In: FEMS Microbiology Letters, Vol. 257, No. 2, 01.04.2006, p. 195-201.

Research output: Contribution to journalArticle

Gao, Guanjun ; Le, Donghai ; Huang, Lifen ; Lu, Huiming ; Narumi, Issay ; Hua, Yuejin. / Internal promoter characterization and expression of the Deinococcus radiodurans pprI-folP gene cluster. In: FEMS Microbiology Letters. 2006 ; Vol. 257, No. 2. pp. 195-201.
@article{82431668914d4630b28346d6d1a481e7,
title = "Internal promoter characterization and expression of the Deinococcus radiodurans pprI-folP gene cluster",
abstract = "PprI is a general gene switch responsible for the extraordinary radioresistance of Deinococcus radiodurans. From NCBI DNA sequence analysis, it was predicted that the translation start codon of the downstream folP (DR0168) gene overlaps the pprI (DR0167) stop codon, suggesting that these genes may form an operon. In this study, we show that a mutant containing an inserted sequence in folP does not grow unless folate is added to the medium, but is not affected in extreme radioresistance, whereas a pprI disruptant strain could grow in the absence of folate. It was found that expression of a pprI-lacZ fusion is constitutive and unaltered following ionizing radiation as is the production of the PprI protein. PprI protein is not expressed if its promoter is deleted and the transcription from the entire pprI promoter is essential for radioresistance of D. radiodurans. However, the deletion of pprI promoter has no effect on the expression of the folP-lacZ fusion. Primer extension analysis of the folP promoter region shows that folP is transcribed from its own promoter located within the pprI structural gene. All these results do support neither the existence of a pprI-folP operon nor a regulatory role of FolP in pprI expression.",
keywords = "Deincoccus radiodurans, folP, lacZ-fusion, pprI, Promoter, Western blot",
author = "Guanjun Gao and Donghai Le and Lifen Huang and Huiming Lu and Issay Narumi and Yuejin Hua",
year = "2006",
month = "4",
day = "1",
doi = "10.1111/j.1574-6968.2006.00169.x",
language = "English (US)",
volume = "257",
pages = "195--201",
journal = "FEMS Microbiology Letters",
issn = "0378-1097",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Internal promoter characterization and expression of the Deinococcus radiodurans pprI-folP gene cluster

AU - Gao, Guanjun

AU - Le, Donghai

AU - Huang, Lifen

AU - Lu, Huiming

AU - Narumi, Issay

AU - Hua, Yuejin

PY - 2006/4/1

Y1 - 2006/4/1

N2 - PprI is a general gene switch responsible for the extraordinary radioresistance of Deinococcus radiodurans. From NCBI DNA sequence analysis, it was predicted that the translation start codon of the downstream folP (DR0168) gene overlaps the pprI (DR0167) stop codon, suggesting that these genes may form an operon. In this study, we show that a mutant containing an inserted sequence in folP does not grow unless folate is added to the medium, but is not affected in extreme radioresistance, whereas a pprI disruptant strain could grow in the absence of folate. It was found that expression of a pprI-lacZ fusion is constitutive and unaltered following ionizing radiation as is the production of the PprI protein. PprI protein is not expressed if its promoter is deleted and the transcription from the entire pprI promoter is essential for radioresistance of D. radiodurans. However, the deletion of pprI promoter has no effect on the expression of the folP-lacZ fusion. Primer extension analysis of the folP promoter region shows that folP is transcribed from its own promoter located within the pprI structural gene. All these results do support neither the existence of a pprI-folP operon nor a regulatory role of FolP in pprI expression.

AB - PprI is a general gene switch responsible for the extraordinary radioresistance of Deinococcus radiodurans. From NCBI DNA sequence analysis, it was predicted that the translation start codon of the downstream folP (DR0168) gene overlaps the pprI (DR0167) stop codon, suggesting that these genes may form an operon. In this study, we show that a mutant containing an inserted sequence in folP does not grow unless folate is added to the medium, but is not affected in extreme radioresistance, whereas a pprI disruptant strain could grow in the absence of folate. It was found that expression of a pprI-lacZ fusion is constitutive and unaltered following ionizing radiation as is the production of the PprI protein. PprI protein is not expressed if its promoter is deleted and the transcription from the entire pprI promoter is essential for radioresistance of D. radiodurans. However, the deletion of pprI promoter has no effect on the expression of the folP-lacZ fusion. Primer extension analysis of the folP promoter region shows that folP is transcribed from its own promoter located within the pprI structural gene. All these results do support neither the existence of a pprI-folP operon nor a regulatory role of FolP in pprI expression.

KW - Deincoccus radiodurans

KW - folP

KW - lacZ-fusion

KW - pprI

KW - Promoter

KW - Western blot

UR - http://www.scopus.com/inward/record.url?scp=33645238983&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645238983&partnerID=8YFLogxK

U2 - 10.1111/j.1574-6968.2006.00169.x

DO - 10.1111/j.1574-6968.2006.00169.x

M3 - Article

C2 - 16553853

AN - SCOPUS:33645238983

VL - 257

SP - 195

EP - 201

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

SN - 0378-1097

IS - 2

ER -