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Plant Cell, Vol. 10, 859-872, May 1998, Copyright © 1998, American Society of Plant Physiologists

Molecular Characterization of the PEND Protein, a Novel bZIP Protein Present in the Envelope Membrane That Is the Site of Nucleoid Replication in Developing Plastids

Naoki Satoa, Kimihisa Ohshimab, Ai Watanabea, Niji Ohtac, Yoshitaka Nishiyamad, Jacques Joyarde, and Roland Doucee
a Department of Biochemistry and Molecular Biology, Faculty of Science, Saitama University, Urawa 338, Japan
b Saitama Prefectural Shim-Misato Water Filtration Plant, 1 Minami-hasunuma, Misato 341, Japan
c School of Human Science, Waseda University, Mikajima, Tokorozawa 359, Japan
d Department of Regulation Biology, National Institute for Basic Biology, Okazaki 444, Japan
e Physiologie Cellulaire Végétale, URA 576 CEA/CNRS/Université Joseph Fourier, Département de Biologie Moléculaire et Structurale, CEA-Grenoble 38054, Grenoble-cedex 9, France

Correspondence to: Naoki Sato, naokisat{at}molbiol.saitama-u.ac.jp (E-mail), 81-48-858-3384 (fax).

Plastid nucleoids are known to bind to the envelope membrane in developing chloroplasts. Here, plastid DNA is extensively replicated. We previously detected a DNA binding protein in the inner envelope membranes of developing plastids in pea and named it PEND (for plastid envelope DNA binding) protein. In this study, we report on the structure and molecular characterization of a cDNA for the PEND protein. As a result of screening cDNA libraries in {lambda}gt11 with one of the target sequences of the PEND protein as a probe, we obtained a clone (PD2) for a novel DNA binding protein consisting of 633 amino acid residues. Analysis of the N-terminal sequence of the purified PEND protein indicated that the transit peptide is just 16 residues long. The PEND protein was detected specifically in the plastid envelope membrane of young unopened leaf buds by immunoblot analysis. The PEND protein consists of a basic region plus zipper region, an unprecedented sextuple repeat region, and a putative membrane-spanning region. The basic region with a zipper region seems to have diverged from that of other plant transcription factors. In addition, the PEND protein could be a distant homolog of the trans-Golgi network integral membrane proteins. The PEND protein is therefore a novel type of DNA binding protein that binds to the membrane as an intrinsic membrane protein.




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