title: Identification of genes required for hydrogenase activity in Chlamydomonas reinhardtii.
abstract: The eukaryotic green alga, Chlamydomonas reinhardtii, produces H(2) under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H(2) production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H(2)-sensor films for clones defective in H(2) production. Two mutants of particular interest were fully characterized. One mutant, hydEF-1, is unable to assemble an active [FeFe]-hydrogenase. This is the first reported C. reinhardtii mutant that is not capable of producing any H(2). The second mutant, sta7-10, is not able to accumulate insoluble starch and has significantly lowered H(2)-photoproduction rates in comparison with the wild-type. In hydEF-1, anaerobiosis induces transcription of the two reported C. reinhardtii hydrogenase genes, HydA1 and HydA2, indicating a normal transcriptional response to anaerobiosis. In contrast, the transcription of both hydrogenase genes in sta7-10 is significantly attenuated.
authors.affiliation: National Renewable Energy Laboratory, 1617 Cole Blvd, Golden, CO 80401, USA.
authors.forename: M C
authors.initials: MC
authors.lastname: Posewitz
authors.forename: P W
authors.initials: PW
authors.lastname: King
authors.forename: S L
authors.initials: SL
authors.lastname: Smolinski
authors.forename: R Davis
authors.initials: RD
authors.lastname: Smith
authors.forename: A R
authors.initials: AR
authors.lastname: Ginley
authors.forename: M L
authors.initials: ML
authors.lastname: Ghirardi
authors.forename: M
authors.initials: M
authors.lastname: Seibert
mesh_terms: D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D006864:Hydrogenase / Q000235:genetics*; D016296:Mutagenesis
chemical_list: D006864:Hydrogenase