In vivo analysis of the roles of conserved aspartate and histidine residues within a complex response regulator.
RcaC is the founding member of a group of large response regulators with complex domain combinations containing at least two receiver domains, an OmpR-class winged helix-turn-helix DNA binding domain, and a histidine phosphotransfer (HPt) domain. Within its two receiver and HPt domains, RcaC contains consensus phosphorylation sites at aspartates 51, 576 and histidine 316. RcaC operates in the pathway regulating transcription of genes encoding components of photosynthetic light harvesting antenna to changes in light colour. We show that phycocyanin gene expression requires RcaC. RcaC contributes to light regulation of phycoerythrin genes, but is not part of the second light regulation pathway controlling these genes. Substitutions at aspartate 51 or histidine 316 severely impaired light responsiveness while substitutions at aspartate 576 had little effect. Complete loss of light regulation, measured by phycocyanin gene expression, only occurred in the triple mutant. We conclude that aspartate 51 primarily controls light colour responsiveness and is regulated by histidine 316, and that these residues are likely phosphorylated in red light and dephosphorylated in green light. The carboxy-terminal receiver domain has a minor role in controlling this response. RcaC abundance is also light regulated and depends on aspartate 51 and histidine 316, but not aspartate 576.
Li
Lina
L
Department of Biology, 1001 East 3rd Street, Indiana University, Bloomington, IN 47405, USA.
Kehoe
David M
DM
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001224:Aspartic Acid / Q000502:physiology*; D017124:Conserved Sequence; D000458:Cyanobacteria / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D006639:Histidine / Q000502:physiology*; D008027:Light; D045342:Light-Harvesting Protein Complexes; D010766:Phosphorylation; D010799:Phycoerythrin / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D015398:Signal Transduction
D045342:Light-Harvesting Protein Complexes; D010799:Phycoerythrin; D001224:Aspartic Acid; D006639:Histidine; D011494:Protein Kinases