ABSTRACT
We describe two related manganese-binding polypeptides with L-arginine metabolizing enzyme activity that can be detected as distinct components (designated PsbY-A1 and PsbY-A2, previously called L-AME) in membranes containing Photosystem II (PS II) from spinach. The polypeptides are bitopic and appear to exist in a heterodimeric form, but only in the chlorophyll a/b lineage of plants. Both proteins are encoded in the nucleus. In spinach and in Arabidopsis thaliana they are both derived from a single-copy gene (psbY) that is translated into a precursor polyprotein of approximately 20 kDa. The processing of the polyprotein is complex and includes at least four cleavage steps. Both polypeptides are exposed N-terminally to the lumenal and C-terminally to the stromal face of the thylakoid membrane.
Subject(s)
Arabidopsis Proteins , Brassicaceae/genetics , Manganese/metabolism , Membrane Proteins/genetics , Plant Proteins , Ureohydrolases/genetics , Amino Acid Sequence , Arabidopsis/genetics , Arginine/metabolism , Base Sequence , Biological Transport , Cell Compartmentation , Cell Nucleus/genetics , DNA, Complementary/genetics , Dimerization , Evolution, Molecular , Gene Dosage , Gene Library , Membrane Proteins/isolation & purification , Membrane Proteins/metabolism , Molecular Sequence Data , Protein Precursors/metabolism , Protein Processing, Post-Translational , Sequence Analysis , Sequence Homology, Amino Acid , Spinacia oleracea/genetics , Ureohydrolases/isolation & purification , Ureohydrolases/metabolismABSTRACT
The promoter of the gene for the subunit III of photosystem I reaction center (psaF) from spinach has been dissected and studied via promoter/GUS gene fusions in transgenic tobacco. It possesses an architecture that differs from any other spinach promoter of genes encoding proteins involved in photosynthesis studied to date. A 42 bp region located between -220 and -179 bp upstream of the transcription start site has been identified that is indispensable for expression and binds a trans-acting factor. Maximal light-response is obtained with a -220/+ 163 bp segment, whereas longer promoter sequences are significantly less effective, indicating the existence of upstream elements with silencer characteristics. F1 seedlings show different spatial expression patterns in darkness or light. Etiolated seedlings display high GUS activity in the upper hypocotyl, the hook region and the vascular tissue of the cotyledons, whereas in light-grown seedlings no activity was detected in the hypocotyl and almost all cells of the cotyledons express the GUS gene.
