Abstract
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 lambdagt11 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.
Full Text
The Full Text of this article is available as a PDF (589.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Claros M. G., von Heijne G. TopPred II: an improved software for membrane protein structure predictions. Comput Appl Biosci. 1994 Dec;10(6):685–686. doi: 10.1093/bioinformatics/10.6.685. [DOI] [PubMed] [Google Scholar]
- Dreses-Werringloer U., Fischer K., Wachter E., Link T. A., Flügge U. I. cDNA sequence and deduced amino acid sequence of the precursor of the 37-kDa inner envelope membrane polypeptide from spinach chloroplasts. Its transit peptide contains an amphiphilic alpha-helix as the only detectable structural element. Eur J Biochem. 1991 Jan 30;195(2):361–368. doi: 10.1111/j.1432-1033.1991.tb15714.x. [DOI] [PubMed] [Google Scholar]
- Garner J., Crooke E. Membrane regulation of the chromosomal replication activity of E. coli DnaA requires a discrete site on the protein. EMBO J. 1996 Jul 1;15(13):3477–3485. [PMC free article] [PubMed] [Google Scholar]
- Herrmann R. G., Kowallik K. V. Multiple amounts of DNA related to the size of chloroplasts. II. Comparison of electron-microscopic and autoradiographic data. Protoplasma. 1970;69(3):365–372. doi: 10.1007/BF01320301. [DOI] [PubMed] [Google Scholar]
- Kaneko T., Sato S., Kotani H., Tanaka A., Asamizu E., Nakamura Y., Miyajima N., Hirosawa M., Sugiura M., Sasamoto S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 1996 Jun 30;3(3):109–136. doi: 10.1093/dnares/3.3.109. [DOI] [PubMed] [Google Scholar]
- Keller W., König P., Richmond T. J. Crystal structure of a bZIP/DNA complex at 2.2 A: determinants of DNA specific recognition. J Mol Biol. 1995 Dec 8;254(4):657–667. doi: 10.1006/jmbi.1995.0645. [DOI] [PubMed] [Google Scholar]
- Menkens A. E., Schindler U., Cashmore A. R. The G-box: a ubiquitous regulatory DNA element in plants bound by the GBF family of bZIP proteins. Trends Biochem Sci. 1995 Dec;20(12):506–510. doi: 10.1016/s0968-0004(00)89118-5. [DOI] [PubMed] [Google Scholar]
- Razin S. V., Gromova I. I., Iarovaia O. V. Specificity and functional significance of DNA interaction with the nuclear matrix: new approaches to clarify the old questions. Int Rev Cytol. 1995;162B:405–448. doi: 10.1016/s0074-7696(08)62623-6. [DOI] [PubMed] [Google Scholar]
- Reeves R., Nissen M. S. The A.T-DNA-binding domain of mammalian high mobility group I chromosomal proteins. A novel peptide motif for recognizing DNA structure. J Biol Chem. 1990 May 25;265(15):8573–8582. [PubMed] [Google Scholar]
- Sato N. A family of cold-regulated RNA-binding protein genes in the cyanobacterium Anabaena variabilis M3. Nucleic Acids Res. 1995 Jun 25;23(12):2161–2167. doi: 10.1093/nar/23.12.2161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato N., Albrieux C., Joyard J., Douce R., Kuroiwa T. Detection and characterization of a plastid envelope DNA-binding protein which may anchor plastid nucleoids. EMBO J. 1993 Feb;12(2):555–561. doi: 10.1002/j.1460-2075.1993.tb05687.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Slater S., Wold S., Lu M., Boye E., Skarstad K., Kleckner N. E. coli SeqA protein binds oriC in two different methyl-modulated reactions appropriate to its roles in DNA replication initiation and origin sequestration. Cell. 1995 Sep 22;82(6):927–936. doi: 10.1016/0092-8674(95)90272-4. [DOI] [PubMed] [Google Scholar]
- Sugiura M. The chloroplast genome. Plant Mol Biol. 1992 May;19(1):149–168. doi: 10.1007/BF00015612. [DOI] [PubMed] [Google Scholar]
- Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zerges W., Rochaix J. D. Low density membranes are associated with RNA-binding proteins and thylakoids in the chloroplast of Chlamydomonas reinhardtii. J Cell Biol. 1998 Jan 12;140(1):101–110. doi: 10.1083/jcb.140.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]