The HCF136 protein is essential for assembly of the photosystem II reaction center in Arabidopsis thaliana

Hcf136 encodes a hydrophilic protein localized in the lumen of stroma thylakoids. Its mutational inactivation in Arabidopsis thaliana results in a photosystem II (PHII)‐less phenotype. Under standard illumination, PSII is not detectable and the amount of photosystem I (PSI) is reduced, which implies that HCF136p may be required for photosystem biogenesis in general. However, at low light, a comparison of mutants with defects in PSII, PSI, and the cytochrome b6f complex reveals that HCF136p regulates selectively biogenesis of PSII. We demonstrate by in vivo radiolabeling of hcf136 that biogenesis of the reaction center (RC) of PSII is blocked. Gel blot analysis and affinity chromatography of solubilized thylakoid membranes suggest that HCF136p associates with a PSII precomplex containing at least D2 and cytochrome b559. We conclude that HCF136p is essential for assembly of the RC of PSII and discuss its function as a chaperone‐like assembly factor.

[1]  G. Ananyev,et al.  High-resolution kinetic studies of the reassembly of the tetra-manganese cluster of photosynthetic water oxidation: proton equilibrium, cations, and electrostatics. , 1996, Biochemistry.

[2]  K. V. van Wijk,et al.  Co-translational Assembly of the D1 Protein into Photosystem II* , 1999, The Journal of Biological Chemistry.

[3]  P. Westhoff,et al.  HCF164 encodes a thioredoxin-like protein involved in the biogenesis of the cytochrome b(6)f complex in Arabidopsis. , 2001, The Plant cell.

[4]  K. V. van Wijk,et al.  Kinetic Resolution of the Incorporation of the D1 Protein into Photosystem II and Localization of Assembly Intermediates in Thylakoid Membranes of Spinach Chloroplasts (*) , 1996, The Journal of Biological Chemistry.

[5]  Petra Fromme,et al.  Crystal structure of photosystem II from Synechococcus elongatus at 3.8 Å resolution , 2001, Nature.

[6]  A. Barkan,et al.  Nuclear genes required for post-translational steps in the biogenesis of the chloroplast cytochrome b6f complex in maize , 1995, Molecular and General Genetics MGG.

[7]  F. Rabanal,et al.  Proof of principle in a de novo designed protein maquette: an allosterically regulated, charge-activated conformational switch in a tetra-alpha-helix bundle. , 2001, Biochemistry.

[8]  Hoober Jk,et al.  Chlorophyll binding to peptide maquettes containing a retention motif. , 2000 .

[9]  I. Arnold,et al.  ATP Synthase of Yeast Mitochondria , 1999, The Journal of Biological Chemistry.

[10]  M. Mann,et al.  A generic strategy to analyze the spatial organization of multi-protein complexes by cross-linking and mass spectrometry. , 2000, Analytical chemistry.

[11]  L. Eichacker,et al.  Function of a chloroplast SRP in thylakoid protein export. , 2001, Biochimica et biophysica acta.

[12]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.

[13]  L. Eichacker,et al.  Assembly of the D1 Precursor in Monomeric Photosystem II Reaction Center Precomplexes Precedes Chlorophyll a–Triggered Accumulation of Reaction Center II in Barley Etioplasts , 1999, Plant Cell.

[14]  A. P. Sane,et al.  The Nucleus-Encoded HCF107 Gene of Arabidopsis Provides a Link between Intercistronic RNA Processing and the Accumulation of Translation-Competent psbH Transcripts in Chloroplasts , 2001, The Plant Cell Online.

[15]  E. Aro,et al.  Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. , 1993, Biochimica et biophysica acta.

[16]  H. Pakrasi Genetic analysis of the form and function of photosystem I and photosystem II. , 1995, Annual review of genetics.

[17]  J. Barber,et al.  Photoassembly of the manganese cluster and oxygen evolution from monomeric and dimeric CP47 reaction center photosystem II complexes. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[18]  P. Westhoff,et al.  A nuclear‐encoded protein of prokaryotic origin is essential for the stability of photosystem II in Arabidopsis thaliana , 1998, The EMBO journal.

[19]  H. Schägger,et al.  Analysis of molecular masses and oligomeric states of protein complexes by blue native electrophoresis and isolation of membrane protein complexes by two-dimensional native electrophoresis. , 1994, Analytical biochemistry.

[20]  F. Wollman,et al.  The biogenesis and assembly of photosynthetic proteins in thylakoid membranes1 , 1999, Biochimica et biophysica acta.

[21]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Barber,et al.  Isolation and Characterization of Monomeric and Dimeric CP47-Reaction Center Photosystem II Complexes* , 1998, The Journal of Biological Chemistry.

[23]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[24]  J. Barber,et al.  Isolation and biochemical characterisation of monomeric and dimeric photosystem II complexes from spinach and their relevance to the organisation of photosystem II in vivo. , 1997, European journal of biochemistry.

[25]  W. Neupert,et al.  Biogenesis of Porin of the Outer Mitochondrial Membrane Involves an Import Pathway via Receptors and the General Import Pore of the Tom Complex , 2001, The Journal of cell biology.

[26]  A. Barkan,et al.  Participation of nuclear genes in chloroplast gene expression. , 2000, Biochimie.

[27]  James Barber,et al.  Three-dimensional structure of the plant photosystem II reaction centre at 8 Å resolution , 1998, Nature.