Differential expression of hydrogen uptake (hup) genes in vegetative and symbiotic cells of Rhizobium leguminosarum

Abstract

The genetic determinants responsible for H₂-uptake (hup genes) in Rhizobium leguminosarum are organized in six transcriptional units, designated regions hupI to hupVI, with region hupI coding for the hydrogenase structural genes (Leyva et al. 1990). Regulation of the expression of hup genes from R. leguminosarum was examined by using hup-lacZ fusions and mRNA dot-blot analysis. None of the six hup regions is transcribed in vegetative cells grown under normal aerobic conditions, whereas all six regions are transcribed in pea bacteroids. Additionally, exposure of cell cultures to low oxygen tensions specifically induces the expression of regions hupV and hupVI. By studying the expression of hupV- and hupVI-lacZ fusions in R. meliloti mutants it was determined that the microaerobic induction of these two regions is dependent on the regulatory fixLJ system, and that this control is exerted through fixK. Such expression was also shown to be nifA and ntrA independent. The functions of the hupV and hupVI gene products are unknown. The possibility that they play a regulatory role in hup gene expression is unlikely, since pea bacteroids from R. leguminosarum Hup⁻ mutants carrying Tn⁵ insertions in regions hupV and hupVI contained normal levels of mRNA transcripts corresponding to the remaining hup regions.