Genes encoding the NAD-reducing hydrogenase of Rhodococcus opacus MR11

被引:38
作者
Grzeszik, C [1 ]
Lubbers, M [1 ]
Reh, M [1 ]
Schlegel, HG [1 ]
机构
[1] UNIV GOTTINGEN,INST MIKROBIOL,D-37077 GOTTINGEN,GERMANY
来源
MICROBIOLOGY-UK | 1997年 / 143卷
关键词
Rhodococcus opacus MR11; NAD-reducing hydrogenase; hox genes;
D O I
10.1099/00221287-143-4-1271
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The dissociation of the soluble NAD-reducing hydrogenase of Rhodococcus opacus MR11 into two dimeric proteins with different catalytic activities and cofactor composition is unique among the NAD-reducing hydrogenases studied so far. The genes of the soluble hydrogenase were localized on a 7.4 kbp Asnl fragment of the linear plasmid pHG201 via heterologous hybridization. Analysis of the nucleotide sequence of this fragment revealed the seven open reading frames ORF1, hoxF, -U, -Y, -H, -W and ORF7. The six latter ORFs belong to the gene cluster of the soluble hydrogenase. Their gene products are highly homologous to those of the NAD-reducing enzyme of Alcaligenes eutrophus H16. The genes hoxF, -U, -Y and -H encode the subunits alpha, gamma, delta and beta, respectively. The gene hoxW encodes a putative protease, which may be essential for C-terminal processing of the beta subunit. Finally, ORF7 encodes a protein which has similarities to cAMP- and cCMP-binding protein kinases, but its function is not known. ORF1, which lies upstream of the hydrogenase gene cluster, encodes a putative transposase found in IS elements of other bacteria. Northern hybridizations and primer extensions using total RNA of autotrophically and heterotrophically grown cells of R. opacus MR11 indicated that the hydrogenase genes are under control of a sigma(70)-like promoter located at the right end of ORF1 and are even transcribed under heterotrophic conditions at a low revel. Furthermore, this promoter was shown to be active in the recombinant Escherichia coli strain LHY1 harbouring the 7.4 kbp Asnl fragment, resulting in overexpression of the hydrogenase genes. Although all four subunits of the soluble hydrogenase were shown via Western immunoblots to be synthesized in E. coli, no active enzyme was detectable.
引用
收藏
页码:1271 / 1286
页数:16
相关论文
共 74 条
  • [1] STUDIES ON A GRAM-POSITIVE HYDROGEN BACTERIUM, NOCARDIA-OPACA STRAIN 1B .1. DESCRIPTION AND PHYSIOLOGICAL CHARACTERIZATION
    AGGAG, M
    SCHLEGEL, HG
    [J]. ARCHIV FUR MIKROBIOLOGIE, 1973, 88 (04): : 299 - 318
  • [2] NICKEL HYDROGENASES - IN SEARCH OF THE ACTIVE-SITE
    ALBRACHT, SPJ
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 1994, 1188 (03): : 167 - 204
  • [3] ALBRACHT SPJ, 1995, BIOSPEKTRUM, V1, P42
  • [4] CHROMOSOMAL LOCALIZATION OF THE HUMAN GENE ENCODING THE 51-KDA SUBUNIT OF MITOCHONDRIAL COMPLEX-I (NDUFV1) TO 11Q13
    ALI, ST
    DUNCAN, AMV
    SCHAPPERT, K
    HENG, HHQ
    TSUI, LC
    CHOW, W
    ROBINSON, BH
    [J]. GENOMICS, 1993, 18 (02) : 435 - 439
  • [5] ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
  • [6] Aragno M., 1992, The prokaryotes: a handbook on the biology of bacteria: ecophysiology, isolation, identification, applications, vol. 1., P344
  • [7] MUTANTS DEFECTIVE IN THE ENERGY-CONSERVING NADH DEHYDROGENASE OF SALMONELLA-TYPHIMURIUM IDENTIFIED BY A DECREASE IN ENERGY-DEPENDENT PROTEOLYSIS AFTER CARBON STARVATION
    ARCHER, CD
    WANG, XH
    ELLIOTT, T
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (21) : 9877 - 9881
  • [8] COMPLEMENTARY-DNA SEQUENCES OF THE 24-KDA AND 21-KDA SUBUNITS OF COMPLEX-I FROM NEUROSPORA
    AZEVEDO, JE
    DUARTE, M
    BELO, JA
    WERNER, S
    VIDEIRA, A
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 1994, 1188 (1-2): : 159 - 161
  • [9] A REGULATORY SUBUNIT OF THE CAMP-DEPENDENT PROTEIN-KINASE DOWN-REGULATED IN APLYSIA SENSORY NEURONS DURING LONG-TERM SENSITIZATION
    BERGOLD, PJ
    BEUSHAUSEN, SA
    SACKTOR, TC
    CHELEY, S
    BAYLEY, H
    SCHWARTZ, JH
    [J]. NEURON, 1992, 8 (02) : 387 - 397
  • [10] THE RELATIONSHIP BETWEEN BASE COMPOSITION AND CODON USAGE IN BACTERIAL GENES AND ITS USE FOR THE SIMPLE AND RELIABLE IDENTIFICATION OF PROTEIN-CODING SEQUENCES
    BIBB, MJ
    FINDLAY, PR
    JOHNSON, MW
    [J]. GENE, 1984, 30 (1-3) : 157 - 166