Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

M. C. Posewitz; P. W. King; S. L. Smolinski; R. Davis Smith; A. R. Ginley; M. L. Ghirardi; M. Seibert

doi:10.1042/BST0330102

Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

M. C. Posewitz, P. W. King, S. L. Smolinski, R. Davis Smith, A. R. Ginley, M. L. Ghirardi, M. Seibert

Research output: Contribution to journal › Article › peer-review

81 Scopus Citations

Abstract

The eukaryotic green alga, Chlamydamonas reinhardtii, produces H ₂ under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H₂ production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H₂-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 H₂. 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.

Original language	American English
Pages (from-to)	102-104
Number of pages	3
Journal	Biochemical Society Transactions
Volume	33
Issue number	1
DOIs	https://doi.org/10.1042/BST0330102 https://doi.org/10.1042/BST0330102
State	Published - 2005

NREL Publication Number

NREL/JA-590-38657

Keywords

Chlamydomonas reinhardtii
H photoproduction
Hydrogenase
Insertional mutagenesis
Metalloprotein assembly
Starch

Access to Document

Cite this

@article{4558409c0c0349cbb67146c9957a31f9,

title = "Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii",

abstract = "The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-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 H2. 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.",

keywords = "Chlamydomonas reinhardtii, H photoproduction, Hydrogenase, Insertional mutagenesis, Metalloprotein assembly, Starch",

author = "Posewitz, {M. C.} and King, {P. W.} and Smolinski, {S. L.} and Smith, {R. Davis} and Ginley, {A. R.} and Ghirardi, {M. L.} and M. Seibert",

year = "2005",

doi = "10.1042/BST0330102",

language = "American English",

volume = "33",

pages = "102--104",

journal = "Biochemical Society Transactions",

issn = "0300-5127",

publisher = "Portland Press, Ltd.",

number = "1",

}

TY - JOUR

T1 - Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

AU - Posewitz, M. C.

AU - King, P. W.

AU - Smolinski, S. L.

AU - Smith, R. Davis

AU - Ginley, A. R.

AU - Ghirardi, M. L.

AU - Seibert, M.

PY - 2005

Y1 - 2005

N2 - The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-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 H2. 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.

AB - The eukaryotic green alga, Chlamydamonas reinhardtii, produces H 2 under anaerobic conditions, in a reaction catalysed by an [FeFe]-hydrogenase. To identify genes that influence H2 production in C. reinhardtii, a library of 6000 colonies on agar plates was screened with sensitive chemochromic H2-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 H2. 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.

KW - Chlamydomonas reinhardtii

KW - H photoproduction

KW - Hydrogenase

KW - Insertional mutagenesis

KW - Metalloprotein assembly

KW - Starch

UR - http://www.scopus.com/inward/record.url?scp=14644393653&partnerID=8YFLogxK

U2 - 10.1042/BST0330102

DO - 10.1042/BST0330102

M3 - Article

C2 - 15667277

AN - SCOPUS:14644393653

SN - 0300-5127

VL - 33

SP - 102

EP - 104

JO - Biochemical Society Transactions

JF - Biochemical Society Transactions

IS - 1

ER -

Identification of Genes Required for Hydrogenase Activity in Chlamydomonas reinhardtii

Abstract

NREL Publication Number

Keywords

Access to Document

Other files and links

Fingerprint

Cite this