Organization and expression of Rhizobium meliloti nitrogen fixation genes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 10 ·1983-05-00 ·Pages 3005-9

Corbin D, Barran L, Ditta G

Abstract

The boundaries of a nif gene cluster in Rhizobium meliloti were determined by Tn5 mutagenesis. These genes are clustered within a 14- to 15-kilobase (kb) region that includes the nitrogenase structural genes. Sequences within 10 kb on either side of this region are devoid of symbiotically essential gene function. RNA blot analysis identified a 5- to 6-kb transcript that corresponds to the nitrogenase structural gene operon. The 5' end of this transcript and its polarity were determined by nuclease S1 mapping. The 5' end of another nif transcript was also identified by nuclease S1 mapping. The promoter regions for these two nif transcripts control transcription in divergent directions and are separated by 1.9 kb of symbiotically unessential DNA. One Tn5 insertion within the nitrogenase operon did not create a polar mutation. The implications of this finding and the overall emerging picture of the genetic organization of this nif region are discussed.

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Corbin D

Department of Biology, University of California at San Diego, La Jolla, California 92093.

Barran L

Ditta G

References (25)

25 references, click to expand

Directed transposon Tn5 mutagenesis and complementation analysis of Rhizobium meliloti symbiotic nitrogen fixation genes.
Cell. 1982 Jun;29(2):551-9 PMID: 6288262
Legume root nodule nitrogenase. Purification, properties, and studies on its genetic control.
Biochim Biophys Acta. 1974 Dec 18;371(2):337-51 PMID: 4215454
Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis.
J Bacteriol. 1982 Jan;149(1):114-22 PMID: 6274841
Polarity of Tn5 insertion mutations in Escherichia coli.
J Bacteriol. 1980 May;142(2):439-46 PMID: 6247321
A general method for site-directed mutagenesis in prokaryotes.
Nature. 1981 Jan 1;289(5793):85-8 PMID: 6256652
Heme biosynthesis in Rhizobium. Identification of a cloned gene coding for delta-aminolevulinic acid synthetase from Rhizobium meliloti.
J Biol Chem. 1982 Aug 10;257(15):8724-30 PMID: 7096330
Bacteriophage T4 late promoters: mapping 5' ends of T4 gene 23 mRNAs.
EMBO J. 1982;1(1):107-14 PMID: 6201352
Nucleotide sequence of the R.meliloti nitrogenase reductase (nifH) gene.
Nucleic Acids Res. 1981 Nov 11;9(21):5711-23 PMID: 6273806
Genetics and regulation of nitrogen fixation.
Annu Rev Microbiol. 1981;35:207-35 PMID: 7027900
Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.
Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347-51 PMID: 7012838
Regulation of nitrogen fixation in Klebsiella pneumoniae: isolation and characterization of strains with nif-lac fusions.
J Bacteriol. 1981 Jan;145(1):348-57 PMID: 7007318
Large plasmids of fast-growing rhizobia: homology studies and location of structural nitrogen fixation (nif) genes.
J Bacteriol. 1981 Mar;145(3):1129-36 PMID: 7009569
Genes controlling early and late functions in symbiosis are located on a megaplasmid in Rhizobium meliloti.
Mol Gen Genet. 1981;184(2):326-33 PMID: 6949001
Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid of R. meliloti.
Mol Gen Genet. 1981;184(2):318-25 PMID: 6949000
The biosynthesis of nitrogenase MoFe protein polypeptides in free-living cultures of Rhizobium japonicum.
Biochim Biophys Acta. 1979 Dec 17;565(2):365-78 PMID: 518887
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.
J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide.
Nucleic Acids Res. 1977;4(5):1539-52 PMID: 19730
Overlapping sequences of Klebsiella pneumoniae nifDNA cloned and characterized.
Mol Gen Genet. 1979 Jul 2;174(1):59-66 PMID: 384162
Purification and characterization of the molybdenum-iron protein component of nitrogenase from soybean nodule bacteroids.
J Biol Chem. 1974 Jan 25;249(2):500-8 PMID: 4855615
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
Molecular weights of ribosomal RNA in relation to evolution.
J Mol Biol. 1968 Dec;38(3):355-65 PMID: 5718556
Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids.
Cell. 1977 Nov;12(3):721-32 PMID: 922889
Clustering of nitrogen fixation (nif) genes in Rhizobium meliloti.
J Bacteriol. 1982 Jan;149(1):221-8 PMID: 6274844
Hybridization of RNA to double-stranded DNA: formation of R-loops.
Proc Natl Acad Sci U S A. 1976 Jul;73(7):2294-8 PMID: 781674
Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels.
J Mol Biol. 1977 Feb 15;110(1):119-46 PMID: 845942

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.80.10.3005

Published

1983-05-00

Pages

3005-9

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC393962

External Links

PubMed source DOI: 10.1073/pnas.80.10.3005 PMC Full Text View journal details

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Organization and expression of Rhizobium meliloti nitrogen fixation genes.

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