title: Rapid, ATP-dependent degradation of a truncated D1 protein in the chloroplast.
abstract: The D1 protein constitutes one of the reaction center subunits of photosystem II and turns over rapidly due to photooxidative damage. Here, we studied the degradation of a truncated D1 protein. A plasmid with a precise deletion in the reading frame of the psbA gene encoding D1 was introduced into the chloroplast of Chlamydomonas reinhardtii. A homoplasmic mutant containing the desired gene was able to synthesize the truncated form of the polypeptide, but could not accumulate significant levels of it. As a consequence, other central photosystem II subunits did not assemble within the thylakoid membrane. In vivo pulse-chase experiments showed that the abnormal D1 protein is rapidly degraded in the light. Degradation was delayed in the light in the presence of an uncoupler, or when cells were incubated in the dark. Pulse-chase experiments performed in vitro indicate that an ATP and metal-dependent protease is responsible for the breakdown process. The paper describes the first in vivo and in vitro functional test for ATP-dependent degradation of a defect polypeptide in chloroplasts. The possible involvement of proteases similar to those removing abnormal proteins in prokaryotic organisms is discussed on the basis of proteases recently identified in chloroplasts.
authors.affiliation: Martin-Luther-Universität Halle-Wittenberg, Institut für Pflanzenphysiologie, Halle-Saale, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Preiss
authors.forename: S
authors.initials: S
authors.lastname: Schrader
authors.forename: U
authors.initials: U
authors.lastname: Johanningmeier
mesh_terms: D000255:Adenosine Triphosphate / Q000502:physiology*; D002736:Chloroplasts / Q000378:metabolism*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D045332:Photosystem II Protein Complex; D010940:Plant Proteins / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D020524:Thylakoids / Q000737:chemistry
chemical_list: D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D010940:Plant Proteins; D018749:RNA, Plant; D000255:Adenosine Triphosphate