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PMID: 7715602 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The cycHJKL genes of Rhizobium meliloti involved in cytochrome c biogenesis are required for "respiratory" nitrate reduction ex planta and for nitrogen fixation during symbiosis.

Molecular & general genetics : MGG ·Vol. 247 ·No. 1 ·1995-04-10 ·Pages 39-47

Kereszt A, Slaska-Kiss K, Putnoky P, Banfalvi Z, Kondorosi A

Abstract

We report the genetic and biochemical analysis of Rhizobium meliloti mutants defective in symbiotic nitrogen fixation (Fix-) and "respiratory" nitrate reduction (Rnr-). The mutations were mapped close to the ade-1 and cys-46 chromosomal markers and the mutated locus proved to be identical to the previously described fix-14 locus. By directed Tn5 mutagenesis, a 4.5 kb segment of the chromosome was delimited in which all mutations resulted in Rnr- and Fix- phenotypes. Nucleotide sequence analysis of this region revealed the presence of four open reading frames coding for integral membrane and membrane-anchored proteins. Biochemical analysis of the mutants showed that the four proteins were necessary for the biogenesis of all cellular c-type cytochromes. In agreement with the nomenclature proposed for rhizobial genes involved in the formation of c-type cytochromes, the four genes were designated cycH, cycJ, cycK, and cycL, respectively. The predicted protein product of cycH exhibited a high degree of similarity to the Bradyrhizobium japonicum counterpart, while CycK and CycL shared more than 50% amino acid sequence identity with the Rhodobacter capsulatus Cc11 and Cc12 proteins, respectively. cycJ encodes a novel membrane anchored protein of 150 amino acids. We suggest that this gene cluster codes for (parts of) a multisubunit cytochrome c haem lyase. Moreover, our results indicate that in R. meliloti c-type cytochromes are required for respiratory nitrate reduction ex planta, as well as for symbiotic nitrogen fixation in root nodules.

Related Genes
Fix RmCycH RmCycJ RmCycK RmCycL Rnr cycH cycJ cycK cycL
MeSH Terms
Amino Acid Sequence Bacterial Proteins/biosynthesis,genetics,metabolism Base Sequence Cytochrome c Group/biosynthesis,genetics DNA Mutational Analysis DNA, Bacterial/genetics Electron Transport Electrophoresis, Polyacrylamide Gel Gene Expression Regulation, Bacterial Genes, Bacterial Medicago sativa/metabolism Membrane Proteins/genetics,metabolism Molecular Sequence Data Multienzyme Complexes Mutagenesis, Insertional Nitrate Reductase Nitrate Reductases/genetics Nitrogen Fixation/genetics Restriction Mapping Sequence Alignment Sinorhizobium meliloti/enzymology,genetics Spectrophotometry/methods Symbiosis
Chemicals
Bacterial Proteins CycJ protein, Bacteria CycK protein, Bacteria Cytochrome c Group DNA, Bacterial Membrane Proteins Multienzyme Complexes Nitrate Reductases Nitrate Reductase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kereszt A
Institute of Genetics, Hungarian Academy of Sciences, Szeged.
Slaska-Kiss K
Putnoky P
Banfalvi Z
Kondorosi A
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00425819
Published
1995-04-10
Pages
39-47
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Databases
GENBANK
U00008
External Links
PubMed source DOI: 10.1007/BF00425819
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