Nucleotide sequence and expression of a deep-sea ribulose-1,5-bisphosphate carboxylase gene cloned from a chemoautotrophic bacterial endosymbiont

Jeffrey L. Stein, Margo Haygood, Horst Felbeck

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The gene coding for ribulose-1,5-bisphosphate carboxylase [RuBisCO; 3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] was cloned from a sulfur-oxidizing chemoautotrophic bacterium that resides as an endosymbiont within the gill tissues of Alvinoconcha hessleri, a gastropod inhabiting deep-sea hydrothermal vents. Nucleotide sequence analysis of the cloned fragment demonstrated that the genes encoding the large (RbcL) and small (RbcS) subunits of the symbiont RuBisCO were organized similarly to the RuBisCO operons of free-living photo- and chemoautotrophic prokaryotes. The symbiont rbcL gene shared the highest degree of nucleotide sequence identity with the cyanobacterium Anabaena (69%) while the rbcS nucleotide sequence shared 61% identity with that of the green alga Chlamydomonas reinhardtii. Comparison with a 153-nucleotide partial rbcL sequence from a symbiont of the bivalve Solemya reidi indicated that the two symbiont sequences shared 85% sequence identity at the nucleotide level and 93% at the amino acid level, suggesting a relatively recent common origin. Escherichia coli transformed with a plasmid carrying the RuBisCO operon of the gastropod symbiont in the proper orientation for transcription from the plasmid lac promoter expressed catalytically active RuBisCO. The presence of enzyme activity suggests the proper assembly of the subunits of this deep-sea RuBisCO into the holoenzyme.

Original languageEnglish (US)
Pages (from-to)8850-8854
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number22
StatePublished - Nov 1990
Externally publishedYes

Fingerprint

Hydrothermal Vents
Oceans and Seas
Gastropoda
Operon
Plasmids
Nucleotides
Anabaena
Genes
Ribulose-Bisphosphate Carboxylase
Chlamydomonas reinhardtii
Holoenzymes
Chlorophyta
Bivalvia
Cyanobacteria
Sulfur
Sequence Analysis
Escherichia coli
Bacteria
Amino Acids
Enzymes

Keywords

  • Alvinoconcha hessleri
  • CO fixation
  • Hydrothermal vents
  • Symbiosis
  • Thiobacilli

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Nucleotide sequence and expression of a deep-sea ribulose-1,5-bisphosphate carboxylase gene cloned from a chemoautotrophic bacterial endosymbiont. / Stein, Jeffrey L.; Haygood, Margo; Felbeck, Horst.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 22, 11.1990, p. 8850-8854.

Research output: Contribution to journalArticle

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abstract = "The gene coding for ribulose-1,5-bisphosphate carboxylase [RuBisCO; 3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] was cloned from a sulfur-oxidizing chemoautotrophic bacterium that resides as an endosymbiont within the gill tissues of Alvinoconcha hessleri, a gastropod inhabiting deep-sea hydrothermal vents. Nucleotide sequence analysis of the cloned fragment demonstrated that the genes encoding the large (RbcL) and small (RbcS) subunits of the symbiont RuBisCO were organized similarly to the RuBisCO operons of free-living photo- and chemoautotrophic prokaryotes. The symbiont rbcL gene shared the highest degree of nucleotide sequence identity with the cyanobacterium Anabaena (69{\%}) while the rbcS nucleotide sequence shared 61{\%} identity with that of the green alga Chlamydomonas reinhardtii. Comparison with a 153-nucleotide partial rbcL sequence from a symbiont of the bivalve Solemya reidi indicated that the two symbiont sequences shared 85{\%} sequence identity at the nucleotide level and 93{\%} at the amino acid level, suggesting a relatively recent common origin. Escherichia coli transformed with a plasmid carrying the RuBisCO operon of the gastropod symbiont in the proper orientation for transcription from the plasmid lac promoter expressed catalytically active RuBisCO. The presence of enzyme activity suggests the proper assembly of the subunits of this deep-sea RuBisCO into the holoenzyme.",
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