Structure and transcription of genes within the β-hbd-adh1 region of Clostridium acetobutylicum P262

Youngleson, Jonathan S.; Lin, Fu-Pang; Reid, Sharon J.; Woods, David R.

doi:10.1016/0378-1097(94)00496-e

Cited by 7 publications

(7 citation statements)

References 7 publications

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“…Because of these homologies, structural and mechanistic similarities would be expected. Interestingly, the nucleotide sequence of etfA also exhibited close homology with that of the fixB gene located upstream of hbd in C. acetobutylicum P262 (71). Judging from the gene structure and arrangement, it is probable that the two are homologous genes from different clostridial strains.…”

Section: Discussionmentioning

confidence: 98%

“…Judging from the gene structure and arrangement, it is probable that the two are homologous genes from different clostridial strains. There are two reasons for the gene designation etfAB (C. acetobutylicum ATCC 824) instead of fixBA (C. acetobutylicum P262 [71]). First, ETF from P. denitrificans has been purified and characterized (29,62) and its encoding gene has been cloned and expressed (3).…”

Section: Discussionmentioning

confidence: 99%

“…There was no transcription termination sequence between any two of these three genes (70,71). In C. difficile, hbd is downstream of a putative crotonase gene and upstream of a thiolase gene, with a termination sequence between the crotonase and hbd genes but no such sequence between hbd and the gene for thiolase (47).…”

Section: Discussionmentioning

confidence: 99%

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Cloning, sequencing, and expression of clustered genes encoding beta-hydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase, and butyryl-CoA dehydrogenase from Clostridium acetobutylicum ATCC 824

1996

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The enzymes ␤-hydroxybutyryl-coenzyme A (CoA) dehydrogenase (BHBD), crotonase, and butyryl-CoA dehydrogenase (BCD) from Clostridium acetobutylicum are responsible for the formation of butyryl-CoA from acetoacetyl-CoA. These enzymes are essential to both acid formation and solvent formation by clostridia. Clustered genes encoding BHBD, crotonase, BCD, and putative electron transfer flavoprotein ␣ and ␤ subunits have been cloned and sequenced. The nucleotide sequence of the crt gene indicates that it encodes crotonase, a protein with 261 amino acid residues and a calculated molecular mass of 28.2 kDa; the hbd gene encodes BHBD, with 282 residues and a molecular mass of 30.5 kDa. Three open reading frames (bcd, etfB, and etfA) are located between crt and hbd. The nucleotide sequence of bcd indicates that it encodes BCD, which consists of 379 amino acid residues and has high levels of homology with various acyl-CoA dehydrogenases. Open reading frames etfB and etfA, located downstream of bcd, encode 27.2-and 36.1-kDa proteins, respectively, and show homology with the fixAB genes and the ␣ and ␤ subunits of the electron transfer flavoprotein. These findings suggest that BCD in clostridia might interact with the electron transfer flavoprotein in its redox function. Primer extension analysis identified a promoter consensus sequence upstream of the crt gene, suggesting that the clustered genes are transcribed as a transcriptional unit and form a BCS (butyryl-CoA synthesis) operon. A DNA fragment containing the entire BCS operon was subcloned into an Escherichia coli-C. acetobutylicum shuttle vector. Enzyme activity assays showed that crotonase and BHBD were highly overproduced in cell extracts from E. coli harboring the subclone. In C. acetobutylicum harboring the subclone, the activities of the enzymes crotonase, BHBD, and BCD were elevated.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

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Cloning, sequencing, and expression of clustered genes encoding beta-hydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase, and butyryl-CoA dehydrogenase from Clostridium acetobutylicum ATCC 824

1996

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“…The adhl gene encodes a NADPH-dependent alcohol dehydrogenase (ADH) with activity for butanol and ethanol [18]. The fixB gene encodes a putative electron transport flavoprotein [19]. The transcription of these genes was investigated to determine the time of expression and whether they were independently expressed or formed part of an operon.…”

Section: Structure and Transcription Of The C Acetobutylicum Fixb [mentioning

confidence: 99%

Regulation of nitrogen metabolism, starch utilisation and the β-hbd—adh1 gene cluster in Clostridium acetobutylicum

Woods¹

1995

FEMS Microbiology Reviews

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The successful genetic manipulation of Clostridium acetobutylicum for the increased production of solvents will depend on an understanding of gene structure and regulation in the bacterium. The glutamine synthetase (glnA) gene is regulated by antisense RNA, transcribed from a downstream promoter, in the opposite direction to the glnA gene. An open reading frame (ORF) was detected downstream of the glnA gene, which has sequence homology to response regulators with anti-termination activity and may be involved in sensing nitrogen conditions. The expression of the linked beta-hbd, adh1 and fixB genes was investigated throughout the bacterial growth cycle by RNA hybridisation techniques. The adh1 gene was independently expressed as a 2.4-kb transcript which peaked at 12 h, immediately prior to the solventogenic phase. The beta-hbd and fixB genes were transcribed throughout the acidogenic and solventogenic phases. A regulator gene, regA, which complements a Bacillus subtilis ccpA mutant, has been identified and sequenced from C. acetobutylicum P262. The regA gene repressed the degradation of starch by an uncharacterised C. acetobutylicum gene, and may therefore play a role in the utilisation of carbohydrate substrates in this organism.

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“…Although this new type of ADH lacked homology with any other previously known ADHs [29], they were demonstrated to utilize ethanol, as well as number of other alcohols, as substrate(s) [6]. In the intervening years, a number of additional microbial enzymes of this iron-activated enzyme type have been described [11,32,50,54,73,83]. Despite the nomenclature of "iron-activated" ADHs, these enzymes are variously activated by a range of divalent cations.…”

Section: Introductionmentioning

confidence: 97%

Differentiation-dependent expression of Adhfe1 in adipogenesis

Kim¹,

Tillison²,

Zhou³

et al. 2007

Archives of Biochemistry and Biophysics

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We have determined that adipocytes are a major site of expression of the transcript for the novel alcohol dehydrogenase (ADH), Adhfe1. Adhfe1 is unique in that the sequence of its encoded protein places it among the iron-activated ADHs. Western blot analysis reveals Adhfe1 encodes a 50 kDa cytoplasmic protein and immunocytochemical staining indicates mitochondrial localization. Adhfe1 transcript exhibits differentiation-dependent expression during in vitro brown and white adipogenesis. Unlike many adipocyte-enriched genes, however, Adhfe1 transcript expression in adipocytes is refractory to TNFα-mediated downregulation. However, use of pharmacological inhibitors reveals PI 3-kinase-mediated signals maintain the basal level of Adhfe1 transcript in 3T3-L1 adipocytes. Tissue profiling studies show Adhfe1 transcript is restricted to white and brown adipose tissues, liver, and kidney. In comparison to C57BL/6 mice, Adhfe1 transcript is downregulated 40% in white adipose tissue of ob/ob obese mice. Further characterization of Adhfe1 should yield new insights into adipocyte function and energy metabolism.

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Structure and transcription of genes within the β-hbd-adh1 region of Clostridium acetobutylicum P262

Cited by 7 publications

References 7 publications

Cloning, sequencing, and expression of clustered genes encoding beta-hydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase, and butyryl-CoA dehydrogenase from Clostridium acetobutylicum ATCC 824

Cloning, sequencing, and expression of clustered genes encoding beta-hydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase, and butyryl-CoA dehydrogenase from Clostridium acetobutylicum ATCC 824

Regulation of nitrogen metabolism, starch utilisation and the β-hbd—adh1 gene cluster in Clostridium acetobutylicum

Differentiation-dependent expression of Adhfe1 in adipogenesis

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