title: Chloroplast His-to-Asp signal transduction: a potential mechanism for plastid gene regulation in Heterosigma akashiwo (Raphidophyceae).
abstract: BACKGROUND
  Maintenance of homeostasis requires that an organism perceive selected physical and chemical signals within an informationally dense environment. Functionally, an organism uses a variety of signal transduction arrays to amplify and convert these perceived signals into appropriate gene transcriptional responses. These changes in gene expression serve to modify selective metabolic processes and thus optimize reproductive success. Here we analyze a chloroplast-encoded His-to-Asp signal transduction circuit in the stramenopile Heterosigma akashiwo (Hada) Hada ex Y. Hara et Chihara [syn. H. carterae (Hulburt) F.J.R. Taylor]. The presence, structure and putative function of this protein pair are discussed in the context of their evolutionary homologues.
  
  
  RESULTS
  Bioinformatic analysis of the Heterosigma akashiwo chloroplast genome sequence revealed the presence of a single two-component His-to-Asp (designated Tsg1/Trg1) pair in this stramenopile (golden-brown alga). These data represent the first documentation of a His-to-Asp array in stramenopiles and counter previous reports suggesting that such regulatory proteins are lacking in this taxonomic cluster. Comparison of the 43 kDa H. akashiwo Tsg1 with bacterial sensor kinases showed that the algal protein exhibits a moderately maintained PAS motif in the sensor kinase domain as well as highly conserved H, N, G1 and F motifs within the histidine kinase ATP binding site. Molecular modelling of the 27 kDa H. akashiwo Trg1 regulator protein was consistent with a winged helix-turn-helix identity - a class of proteins that is known to impact gene expression at the level of transcription. The occurrence of Trg1 protein in actively growing H. akashiwo cells was verified by Western analysis. The presence of a PhoB-like RNA polymerase loop in Trg1 and its homologues in the red-algal lineage support the hypothesis that Trg1 and its homologues interact with a sigma 70 (sigma70) subunit (encoded by rpoD) of a eubacterial type polymerase. Sequence analysis of H. akashiwo rpoD showed this nuclear-encoded gene has a well-defined 4.2 domain, a region that augments RNA polymerase interaction with transcriptional regulatory proteins and also serves in -35 promoter recognition. The presence/loss of the His-to-Asp pairs in primary and secondary chloroplast lineages is assessed.
  
  
  CONCLUSIONS
  His-to-Asp signal transduction components are found in most rhodophytic chloroplasts, as well as in their putative cyanobacterial progenitors. The evolutionary conservation of these proteins argues that they are important for the maintenance of chloroplast homeostasis. Our data suggest that chloroplast gene transcription may be impacted by the interaction of the His-to-Asp regulator protein (which is less frequently lost than the sensor protein) with the RNA polymerase sigma70 subunit.
authors.affiliation: Department of Biology, University of Washington, Seattle, WA 98195-5325, USA. mdupless@u.washington.edu <mdupless@u.washington.edu>
authors.forename: Melinda R
authors.initials: MR
authors.lastname: Duplessis
authors.forename: Kenneth G
authors.initials: KG
authors.lastname: Karol
authors.forename: Elinor T
authors.initials: ET
authors.lastname: Adman
authors.forename: Lauren Y S
authors.initials: LY
authors.lastname: Choi
authors.forename: Michael A
authors.initials: MA
authors.lastname: Jacobs
authors.forename: Rose Ann
authors.initials: RA
authors.lastname: Cattolico
mesh_terms: D020418:Algal Proteins / Q000235:genetics*; D001224:Aspartic Acid / Q000235:genetics; D002736:Chloroplasts / Q000235:genetics*; D044523:Chrysophyta / Q000235:genetics*; D005786:Gene Expression Regulation; D006639:Histidine / Q000235:genetics; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D020418:Algal Proteins; D014157:Transcription Factors; D001224:Aspartic Acid; D006639:Histidine