GntR-Type Transcriptional Regulator pckr Negatively Regulates the Expression of Phosphoenolpyruvate Carboxykinase in Corynebacterium glutamicum

Jeong Eun Hyeon, Dae Hee Kang, Young In Kim, Seung Kyou You, Sung Ok Han

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The pck (cg3169) gene of Corynebacterium glutamicum encodes a phosphoenolpyruvate carboxykinase (PEPCK). Here, a candidate transcriptional regulator that binds to the promoter region of pck was detected using a DNA affinity purification approach. An isolated protein was identified to be PckR (Cg0196), a GntR family transcriptional regulator which consists of 253 amino acids with a mass of 27 kDa as measured by peptide mass fingerprinting. The results of electrophoretic mobility shift assays verified that PckR specifically binds to the pck promoter. The putative regulator binding region extended from position-44 to-27 (an 18-bp sequence) relative to the transcriptional start point of the pck gene. We measured the expression of pck in a pckR deletion mutant by using quantitative real-time reverse transcription-PCR. The expression level of pck in the pckR mutant was 7.6 times higher than that in wild-type cells grown in glucose. Comparative DNA microarray hybridizations and bioinformatic searches revealed the gene composition of the transcriptional regulon of C. glutamicum. Based on these results, PckR seemed to play an important role in the regulation of PEPCK in C. glutamicum grown in glucose. In particular, these assays revealed that PckR acts as a repressor of pck expression during glucose metabolism.

Original languageEnglish
Pages (from-to)2181-2188
Number of pages8
JournalJournal of Bacteriology
Volume194
Issue number9
DOIs
Publication statusPublished - 2012 May 1

Fingerprint

Corynebacterium glutamicum
Phosphoenolpyruvate
Glucose
Genes
Regulon
Peptide Mapping
Electrophoretic Mobility Shift Assay
Oligonucleotide Array Sequence Analysis
Computational Biology
Genetic Promoter Regions
Reverse Transcription
Amino Acids
Polymerase Chain Reaction
DNA
Proteins

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

GntR-Type Transcriptional Regulator pckr Negatively Regulates the Expression of Phosphoenolpyruvate Carboxykinase in Corynebacterium glutamicum. / Hyeon, Jeong Eun; Kang, Dae Hee; Kim, Young In; You, Seung Kyou; Han, Sung Ok.

In: Journal of Bacteriology, Vol. 194, No. 9, 01.05.2012, p. 2181-2188.

Research output: Contribution to journalArticle

@article{d0f985ecffaf4d0cb48d9f456b687b6f,
title = "GntR-Type Transcriptional Regulator pckr Negatively Regulates the Expression of Phosphoenolpyruvate Carboxykinase in Corynebacterium glutamicum",
abstract = "The pck (cg3169) gene of Corynebacterium glutamicum encodes a phosphoenolpyruvate carboxykinase (PEPCK). Here, a candidate transcriptional regulator that binds to the promoter region of pck was detected using a DNA affinity purification approach. An isolated protein was identified to be PckR (Cg0196), a GntR family transcriptional regulator which consists of 253 amino acids with a mass of 27 kDa as measured by peptide mass fingerprinting. The results of electrophoretic mobility shift assays verified that PckR specifically binds to the pck promoter. The putative regulator binding region extended from position-44 to-27 (an 18-bp sequence) relative to the transcriptional start point of the pck gene. We measured the expression of pck in a pckR deletion mutant by using quantitative real-time reverse transcription-PCR. The expression level of pck in the pckR mutant was 7.6 times higher than that in wild-type cells grown in glucose. Comparative DNA microarray hybridizations and bioinformatic searches revealed the gene composition of the transcriptional regulon of C. glutamicum. Based on these results, PckR seemed to play an important role in the regulation of PEPCK in C. glutamicum grown in glucose. In particular, these assays revealed that PckR acts as a repressor of pck expression during glucose metabolism.",
author = "Hyeon, {Jeong Eun} and Kang, {Dae Hee} and Kim, {Young In} and You, {Seung Kyou} and Han, {Sung Ok}",
year = "2012",
month = "5",
day = "1",
doi = "10.1128/JB.06562-11",
language = "English",
volume = "194",
pages = "2181--2188",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "9",

}

TY - JOUR

T1 - GntR-Type Transcriptional Regulator pckr Negatively Regulates the Expression of Phosphoenolpyruvate Carboxykinase in Corynebacterium glutamicum

AU - Hyeon, Jeong Eun

AU - Kang, Dae Hee

AU - Kim, Young In

AU - You, Seung Kyou

AU - Han, Sung Ok

PY - 2012/5/1

Y1 - 2012/5/1

N2 - The pck (cg3169) gene of Corynebacterium glutamicum encodes a phosphoenolpyruvate carboxykinase (PEPCK). Here, a candidate transcriptional regulator that binds to the promoter region of pck was detected using a DNA affinity purification approach. An isolated protein was identified to be PckR (Cg0196), a GntR family transcriptional regulator which consists of 253 amino acids with a mass of 27 kDa as measured by peptide mass fingerprinting. The results of electrophoretic mobility shift assays verified that PckR specifically binds to the pck promoter. The putative regulator binding region extended from position-44 to-27 (an 18-bp sequence) relative to the transcriptional start point of the pck gene. We measured the expression of pck in a pckR deletion mutant by using quantitative real-time reverse transcription-PCR. The expression level of pck in the pckR mutant was 7.6 times higher than that in wild-type cells grown in glucose. Comparative DNA microarray hybridizations and bioinformatic searches revealed the gene composition of the transcriptional regulon of C. glutamicum. Based on these results, PckR seemed to play an important role in the regulation of PEPCK in C. glutamicum grown in glucose. In particular, these assays revealed that PckR acts as a repressor of pck expression during glucose metabolism.

AB - The pck (cg3169) gene of Corynebacterium glutamicum encodes a phosphoenolpyruvate carboxykinase (PEPCK). Here, a candidate transcriptional regulator that binds to the promoter region of pck was detected using a DNA affinity purification approach. An isolated protein was identified to be PckR (Cg0196), a GntR family transcriptional regulator which consists of 253 amino acids with a mass of 27 kDa as measured by peptide mass fingerprinting. The results of electrophoretic mobility shift assays verified that PckR specifically binds to the pck promoter. The putative regulator binding region extended from position-44 to-27 (an 18-bp sequence) relative to the transcriptional start point of the pck gene. We measured the expression of pck in a pckR deletion mutant by using quantitative real-time reverse transcription-PCR. The expression level of pck in the pckR mutant was 7.6 times higher than that in wild-type cells grown in glucose. Comparative DNA microarray hybridizations and bioinformatic searches revealed the gene composition of the transcriptional regulon of C. glutamicum. Based on these results, PckR seemed to play an important role in the regulation of PEPCK in C. glutamicum grown in glucose. In particular, these assays revealed that PckR acts as a repressor of pck expression during glucose metabolism.

UR - http://www.scopus.com/inward/record.url?scp=84861210999&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84861210999&partnerID=8YFLogxK

U2 - 10.1128/JB.06562-11

DO - 10.1128/JB.06562-11

M3 - Article

VL - 194

SP - 2181

EP - 2188

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 9

ER -