Journal of Experimental Botany, Vol. 51, No. 343, pp. 317-318,
February 2000
© 2000 Oxford University Press
Gene Note |
A novel Arabidopsis thaliana dynamin-like protein containing the pleckstrin homology domain1
2 Laboratory of Plant Molecular Biology, Institute of Physical and Chemical Research (RIKEN), Tsukuba Life Science Center, 3–1-1 Koyadai, Tsukuba, Ibaraki 305–0074, Japan
3 Biological Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Ministry of Agriculture, Forestry and Fisheries, 1–2 Ohwashi, Tsukuba, Ibaraki 305–8686, Japan
Received 6 July 1999; Accepted 30 September 1999
Abstract
A full-length cDNA encoding a novel type of plant dynamin-like protein, ADL3, was isolated from Arabidopsis thaliana. ADL3 is a high molecular weight GTPase whose GTP-binding domain shows a low homology to those of other plant dynamin-like proteins. ADL3 contains the pleckstrin homology domain as is in mammalian dynamins, although other plant dynamin-like proteins reported lack this domain. The ADL3 gene was expressed weakly in various tissues, except for siliques with high level expression, which is distinct from the case for other plant dynamin-like protein genes. Taken together, it is predicted that the mode of activation of ADL3 is different from those of other plant homologues.
Key words: Arabidopsis thaliana, dynamin-like protein, pleckstrin homology (PH) domain.
Dynamin, the high molecular weight GTPase, plays a key role in scission event common in various type of endocytosis at plasma membrane (van der Bliek, 1999
). Dynamins usually consist of N-terminal GTPase domain, pleckstrin homology (PH) domain and C-terminal proline/arginine-rich domain (PRD) with a conserved domain of unknown function and coiled-coil domain. The PH domain and PRD are well known to have the capacity to exert regulatory effects on the GTPase activity (Lin et al., 1997; Herskovits et al., 1993).
Dynamin-like proteins have been identified in higher plants. It is notable that all plant dynamin-like proteins reported lack the PH domain and PRD. In yeast, a dynamin-like protein, Vps1p, is involved in protein transport from Golgi to an endosomal compartment (Wilsbach and Payne, 1993), while another dynamin-like protein, Mgm1p, plays a role in mitochondria maintenance (Jones and Fangman, 1992). Interestingly, these yeast proteins also lack the PH domain and PRD. It is therefore probable that plant dynamin-like proteins also function in intracellular events other than in endocytosis. Indeed, soybean PDL (plant
dynamin-like) is required for the formation of cell plates from Golgi (Gu and Verma, 1996) and Arabidopsis ADL1 (Aumrabidopsis
dynamin-like
1) functions in biogenesis of thylakoid membranes in chloroplasts (Park et al., 1998). However, the existence of the PH domain in plants was currently confirmed by the molecular cloning of Arabidopsis cDNAs for phosphatidylinositol 4-kinase (PI 4-K) (Stevenson et al., 1998) and pleckstrin homologue (Mikami et al., 1999), which led us to search a plant dynamin-like protein containing the PH domain.
Searching of the plant expression sequence tag (EST) databases against human pleckstrin resulted in the discovery of an Arabidopsis EST clone, 208A13T7 (GenBank accession number N37665), encoding a part of the PH domain. A DNA fragment corresponding to the EST sequence was amplified by polymerase chain reaction (PCR) and used as a probe for screening of an Arabidopsis cDNA library. The largest cDNA insert, 3172 bp, was sequenced, yielding a 2508 bp complete reading frame (from nucleotide positions 117 to 2624) for a protein of 836 amino acids with calculated molecular mass of 91.6 kDa. Based on the structural characteristics (Fig. 1A), a protein encoded by the cDNA was thought to be a member of plant dynamin-like proteins and designated ADL3. The cDNAs for ADL1 and ADL2 have already been isolated from Arabidopsis plants (Dombrowski and Raikhel, 1995; Kang et al., 1998). Southern blotting analysis indicated that ADL3 is encoded by a single copy gene (data not shown).
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-helix in the PH domain are underlined and double-underlined,respectively. (C) Multiple amino acid sequence alignment of humandynamin-1 and Arabidopsis dynamin-like proteins. Residues indicatedin white letters on a black background are identical in 3 to 4 sequencesand those possibly involved in GTP-binding are indicated by asterisks.Accession numbers: ADL1, As shown in Fig. 1A, ADL3 contains the PH domain but not the PRD, although amino acid sequences of the PH domain of ADL3 is not homologous to that of human dynamin-1 (Fig. 1B). The GTP-binding domain of ADL3 shows a low homology to those of other proteins (28% to ADL1, 27% to ADL2 and 29% to human dynamin-1) (Fig. 1C), thus ADL3 is probably different in endogenous GTPase activity and affinity to GTP from other proteins. In addition, the N-terminal extension of ADL3 is not homologous to those of other proteins (Fig. 1C), suggesting the difference in subcellular localization among them. Although the N-terminal extension of ADL2 has been identified as the chloroplast-targeting signal (Kang et al., 1998
), it is difficult to speculate to which ADL3 is sorted, because the prediction of protein localization sites (http://www.nibb.ac.jp:8000) indicate that the N-terminal extension of ADL3 is not typical signal sequence and that ADL3 has a potential to translocate to chloroplast stroma, mitochondrial matrix space, microbody, and nucleus. It is, however, possible that ADL3 associates with membranes via the interaction between its PH domain and phosphoinositides of membranes. Moreover, the ADL3 mRNA was expressed weakly in various tissues, except in the siliques with high level abundance (Fig. 2), which is different from those of ADL1 gene in roots, stems and flowers (Dombrowski and Raikhel, 1995), ADL2 gene in flowers at highest level (Kang et al., 1998) and soybean PDL gene in roots and stems (Gu and Verma, 1996). Based on above findings, it is highly possible that the mode of activation of ADL3 is different from those of other plant dynamin-like proteins and that ADL3 presumably perform a role different from other related proteins. Therefore, functional characterization of ADL3 must provide new insight into the biological significance of dynamin-like proteins in higher plants.
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Acknowledgments
We thank to Dr K Ichimura for his helpful discussion and Mr N Sugai for his technical assistance. This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences to KY-S. and by a Grant-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan and the Special Coordination Fund of the Science and Technology Agency of Japan to KS.
Notes
1 The sequence of the cDNA reported here has been deposited in DDBJ/EMBL/GenBank DNA databases with the accession no. AB026987. 
4 Present address: Department of Regulation Biology, National Institute for Basic Biology, Myodaiji-cho, Okazaki 444–8585, Japan.
5 To whom correspondence should be addressed. Fax: +81 564 54 4866. E-mail:mikami{at}nibb.ac.jp
References
Dombrowski JE, Raikhel NV. 1995. Isolation of a cDNA encoding a novel GTP-binding protein of Arabidopsis thaliana. Plant Molecular Biology 28, 1121–1126.[Web of Science][Medline]
Gu X, Verma DPS. 1996. Phragmoplastin, a dynamin-like protein associated with cell plate formation in plants. EMBO Journal 15, 695–704.[Web of Science][Medline]
Herskovits JS, Shpetner HS, Burgess CC, Vallee RB. 1993. Microtubles and Srk homology 3 domains stimulate the dynamin GTPase via its C-terminal domain. Proceeding of the National Academy of Sciences, USA 90, 11468–11472.
Jones BA, Fangman WL. 1992. Mitochondrial DNA maintenance in yeast requires a protein containing a region related to the GTP-binding domain of dynamin. Genes and Development 6, 380–389.
Kang SG, Jin JB, Piao HL, Pih KT, Jang HJ, Lim JH, Hwang I. 1998. Molecular cloning of an Arabidopsis cDNA encoding a dynamin-like protein that is localized to plastids. Plant Molecular Biology 38, 437–447.[Web of Science][Medline]
Lin HC, Barylko B, Achiriloaie M, Albanesi JP. 1997. Phosphatidylinositol (4,5)-bisphosphate-dependent activation of dynamins I and II lacking the proline/arginine-rich domains. Journal of Biological Chemistry 272, 25999–26004.
Mikami K, Takahashi S, Katagiri T, Yamaguchi-Shinozaki K, Shinozaki K. 1999. Isolation of an Arabidopsis thaliana cDNA encoding a pleckstrin homology domain protein, a putative homologue of human pleckstrin. Journal of Experimental Botany 50, 729–730.
Park JM, Cho JH, Kang SG, Jang HJ, Pih KT, Piao HL, Cho MJ, Hwang I. 1998. A dynamin-like protein in Arabidopsis thaliana is involved in biogenesis of thylakoid membranes. EMBO Journal 17, 859–867.[Web of Science][Medline]
Stevenson JM, Perera IY, Boss WF. 1998. A phosphatidylinositol 4-kinase pleckstrin homology domain that binds phosphatidylinositol 4-monophosphate. Journal of Biological Chemistry 273, 22761–22767.
van der Bliek AM. 1999. Functional diversity in the dynamin family. Trends in Cell Biology 9, 96–102.[Web of Science][Medline]
Wilsbach K, Payne GS. 1993. Vps1p, a member of the dynamin GTPase family, is necessary for Golgi membrane protein retention in Saccharomyces cerevisiae. EMBO Journal 12, 3049–3059.[Web of Science][Medline]
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