The 9-kDa phosphoprotein of photosystem II. Generation and characterisation of Chlamydomonas mutants lacking PSII-H and a site-directed mutant lacking the phosphorylation site

Biochim Biophys Acta. 1998 Apr 14;1364(1):63-72. doi: 10.1016/s0005-2728(98)00013-9.

Abstract

The chloroplast gene psbH encodes a 9-10 kDa thylakoid membrane protein (PSII-H) that is associated with photosystem II and is subject to light-dependent phosphorylation at a threonine residue located on the stromal side of the membrane. The function of PSII-H is not known, neither is it clear what regulatory role phosphorylation may play in the control of PSII activity. Using particle gun-mediated transformation, we have created chloroplast transformants of Chlamydomonas reinhardtii in which the synthesis of PSII-H is prevented by the disruption of psbH, or in which the phosphorylatable threonine is replaced by alanine through site-directed mutagenesis of the gene. The mutants lacking PSII-H have a photosystem II-deficient phenotype, with no detectable functioning PSII complex present in whole cells or isolated thylakoid membranes. In contrast, the alanine mutant (T3A) grows photoautotrophically, and PSII activity is comparable to wild-type cells as determined by various biochemical and biophysical assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlamydomonas reinhardtii / genetics
  • Chlamydomonas reinhardtii / metabolism*
  • Chloroplasts / metabolism
  • DNA Primers
  • Electron Spin Resonance Spectroscopy
  • Kinetics
  • Light
  • Molecular Weight
  • Mutagenesis, Site-Directed
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Phosphorylation
  • Photosynthetic Reaction Center Complex Proteins / biosynthesis
  • Photosynthetic Reaction Center Complex Proteins / chemistry
  • Photosynthetic Reaction Center Complex Proteins / genetics
  • Photosynthetic Reaction Center Complex Proteins / metabolism*
  • Photosystem II Protein Complex*
  • Polymerase Chain Reaction
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Restriction Mapping
  • Threonine

Substances

  • DNA Primers
  • Phosphoproteins
  • Photosynthetic Reaction Center Complex Proteins
  • Photosystem II Protein Complex
  • Recombinant Proteins
  • photosystem II, psbH subunit
  • Threonine