Expression and Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase Genes in Mycobacterium sp. Strain JC1 DSM 3803

Jae Ho Lee; Dong Oh Park; Sae Woong Park; Eun Ha Hwang; Jeong Il Oh; Young Min Kim

doi:10.1007/s12275-008-0210-3

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Research Support, Non-U.S. Gov't
Expression and Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase Genes in Mycobacterium sp. Strain JC1 DSM 3803: Jae Ho Lee ¹, Dong Oh Park ¹, Sae Woong Park ¹, Eun Ha Hwang ¹, Jeong Il Oh ², Young Min Kim ¹; Journal of Microbiology 2009;47(3):297-307.
DOI: https://doi.org/10.1007/s12275-008-0210-3
Published online: June 26, 2009

¹Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea, ²Department of Microbiology, Pusan National University, Busan 609-735, Republic of Korea¹Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea, ²Department of Microbiology, Pusan National University, Busan 609-735, Republic of Korea

Corresponding author: Young Min Kim , Tel: 82-2-2123-2658,

Received: 26 August 2008 • Accepted: 15 March 2009

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Abstract

Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the key enzyme of the Calvin reductive pentose phosphate cycle. Two sets of structural genes (cbbLS-1 and -2) for form I RubisCO have been previously identified in the Mycobacterium sp. strain JC1, which is able to grow on carbon monoxide (CO) or methanol as sole sources of carbon and energy. Northern blot and reverse transcriptase PCR showed that the cbbLS-1 and -2 genes are expressed in cells grown on either carbon monoxide (CO) or methanol, but not in cells grown in nutrient broth. A promoter assay revealed that the cbbLS-2 promoter has a higher activity than the cbbLS-1 promoter in both CO- and methanol-grown cells, and that the activities of both promoters were higher in CO-grown cells than in methanol-grown cells. A gel mobility shift assay and footprinting assays showed that CbbR expressed in Escherichia coli from a cbbR gene, which is located downstream of cbbLS-1 and transcribed in the same orientation as that of the cbbLS genes, specifically bound to the promoter regions of the cbbLS-1 and -2 genes containing inverted repeat sequence. A DNase I footprinting assay revealed that CbbR protected positions -59 to -3 and -119 to -78 of the cbbLS-1 and -2 promoters, respectively. Overexpression of CbbR induced the transcription of RubisCO genes in Mycobacterium sp. strain JC1 grown in nutrient broth. Our results suggest that the CbbR product from a single cbbR gene may positively regulate two cbbLS operons in the Mycobacterium sp. strain JC1 as is the case for Rhodobacter sphaeroides and Cupriavidus necator.

Keywords: RubisCO genes;two copies of cbbLS genes;gene expression and regulation;mycobacteria;carboxydobacteria;methylotrophic bacteria

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Expression and Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase Genes in Mycobacterium sp. Strain JC1 DSM 3803

J. Microbiol. 2009;47(3):297-307. Published online June 26, 2009

DOI: https://doi.org/10.1007/s12275-008-0210-3

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