RETRACTED: Transcriptional analysis of the F0F1 ATPase operon of Corynebacterium glutamicum ATCC 13032 reveals strong induction by alkaline pH (Retracted Article)

被引:24
作者
Barriuso-Iglesias, M
Barreiro, C
Flechoso, F
Martín, JF
机构
[1] INBIOTEC, Leon 24006, Spain
[2] Univ Leon, Fac Ciencias Biol & Ambientales, E-24071 Leon, Spain
来源
MICROBIOLOGY-SGM | 2006年 / 152卷
关键词
D O I
10.1099/mic.0.28383-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Corynebacterium glutamicum, a soil Gram-positive bacterium used for industrial amino acid production, was found to grow optimally at pH 7.0-9.0 when incubated in 5 litre fermenters under pH-controlled conditions. The highest biomass was accumulated at pH 9.0. Growth still occurred at pH 9.5 but at a reduced rate. The expression of the pH-regulated F0F1 ATPase operon (containing the eight genes atpBEFHAGDC) was induced at alkaline pH. A 7.5 kb transcript, corresponding to the eight-gene operon, was optimally expressed at pH 9.0. The same occurred with a 1.2 kb transcript corresponding to the atpB gene. RT-PCR studies confirmed the alkaline pH induction of the F0F1 operon and the existence of the atpl gene. The atpl gene, located upstream of the F0F1 operon, was expressed at a lower level than the polycistronic 7.5 kb mRNA, from a separate promoter (P-atp1). Expression of the major promoter of the F0F1 operon, designated P-atp2, and the P-atp1 promoter was quantified by coupling them to the pET2 promoter-probe vector. Both P-atp1 and P-atp2 were functional in C. glutamicum and Escherichia coli. Primer extension analysis identified one transcription start point inside each of the two promoter regions. The P-atp1 promoter fitted the consensus sequence of promoters recognized by the vegetative sigma factor of C. glutamicum, whereas the -35 and -10 boxes of P-atp2 fitted the consensus sequence for a sigma(H)-recognized Mycobacterium tuberculosis promoters C-C/(G)GG(A)/(G)AC 17-22 nt (C)/(G)GTT(C)/G, known to be involved in expression of heat-shock and other stress-response genes. These results suggest that the F0F1 operon is highly expressed at alkaline pH, probably using a sigma(H) RNA polymerase.
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页码:11 / 21
页数:11
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