Complementation of cobalamin auxotrophy in Synechococcus sp. strain PCC 7002 and validation of a putative cobalamin riboswitch in vivo

Adam A. Pérez, Zhenfeng Liu, Dmitry A. Rodionov, Zhongkui Li, Donald A. Bryant

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The euryhaline cyanobacterium Synechococcus sp. strain PCC 7002 has an obligate requirement for exogenous vitamin B12 (cobalamin), but little is known about the roles of this compound in cyanobacteria. Bioinformatic analyses suggest that only the terminal enzyme in methionine biosynthesis, methionine synthase, requires cobalamin as a coenzyme in Synechococcus sp. strain PCC 7002. Methionine synthase (MetH) catalyzes the transfer of a methyl group from N5-methyl-5,6,7,8-tetrahydrofolate to L-homocysteine during L-methionine synthesis and uses methylcobalamin as an intermediate methyl donor. Numerous bacteria and plants alternatively employ a cobalamin-independent methionine synthase isozyme, MetE, that catalyzes the same methyl transfer reaction as MetH but uses N5-methyl-5,6,7,8-tetrahydrofolate directly as the methyl donor. The cobalamin auxotrophy of Synechococcus sp. strain PCC 7002 was complemented by using the metE gene from the closely related cyanobacterium Synechococcus sp. strain PCC 73109, which possesses genes for both methionine synthases. This result suggests that methionine biosynthesis is probably the sole use of cobalamin in Synechococcus sp. strain PCC 7002. Furthermore, a cobalaminrepressible gene expression system was developed in Synechococcus sp. strain PCC 7002 that was used to validate the presence of a cobalamin riboswitch in the promoter region of metE from Synechococcus sp. strain PCC 73109. This riboswitch acts as a cobalamin- dependent transcriptional attenuator for metE in that organism.

Original languageEnglish (US)
Pages (from-to)2743-2752
Number of pages10
JournalJournal of bacteriology
Volume198
Issue number19
DOIs
StatePublished - 2016

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

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