Abstract
A cDNA clone encoding a WD-40 repeat protein (BGB1) was characterized in Brassica napus L. The clone contained an open reading frame of 327 amino acid residues almost entirely composed of seven segments of WD-40 repeats. Among the WD-40 repeat proteins, BGB1 showed high similarity (63% identity) to a rat intracellular receptor for protein kinase C (RACK1) that functions in the translocation of activated protein kinase C (PKC) from the cytosolic fraction to the membrane fraction. BGB1 also had two sequence motifs involved in binding of RACK1 to PKC. The cDNA clone, when carried in a Xenopus oocyte expression vector and injected into Xenopus laevis oocytes, inhibited insulin-induced maturation of the oocytes, a PKC-mediated pathway, and this inhibition was accompanied by reduction of PKC in the membrane fraction, as in the case of mammalian RACKs. The data show that BGB1 shares some common functional characteristics with the mammalian RACK1 along with the structural similarity, suggesting that a mammalian RACK1-related cellular process might be operating in plants. Southern blot analyses of the genome of B. napus and Arabidopsis thaliana (L.) Heynh. revealed that BGB1-related genes constitute a small multigene family in both species. An approximately 1.4-kb transcript was constitutively expressed in all organs examined.
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Abbreviations
- CAT:
-
chloramphenicol acetyl transferase
- pOEV:
-
Xenopus oocyte expression vector
- PKC:
-
protein kinase C
- PLC:
-
phospholipase C
- RACK:
-
intracellular receptor for activated protein kinase C
- UTR:
-
untranslated region
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215: 403–310
Deng X-W, Matsui M, Wei N, Wagner D, Chu AM, Feldmann KA, Quail PH (1992) COP1, an Arabidopsis regulatory gene, encodes a protein with both a zinc-binding motif and a Gβ homologous domain. Cell 71: 791–801
Disatnik M-H, Hernandez-Sotomayor SMT, Jones G, Carpenter G, Mochly-Rosen D (1994) Phospholipase C-γl binding to intracellular receptors for activated protein kinase C. Proc Natl Acad Sci USA 91: 559–563
Guillemot F, Billault A, Auffray C (1989) Physical linkage of a guanine nucleotide-binding protein-related gene to the chicken major histocompatibility complex. Proc Natl Acad Sci USA 86: 4594–4598
Han J-K, Lee S-K (1995) Reducing PIP, hydrolysis, Ins(l,4,5)P3 receptor availability, or calcium gradients inhibits progesterone-stimulated Xenopus oocyte maturation. Biochem Biophys Res Commun 217: 931–939
Ishida S, Takahashi Y, Nagata T (1993) Isolation of cDNA of an auxin-regulated gene encoding a G protein β subunit-like protein from tobacco BY-2 cells. Proc Natl Acad Sci USA 90: 11152–11156
Iwasaki Y, Komano M, Ishikawa A, Sasaki T, Asahi T (1995) Molecular cloning and characterization of cDNA for a rice protein that contains seven repetitive segments of the Trp-Asp forty-amino-acid repeat (WD-40 repeat). Plant Cell Physiol 36: 505–510
Kwak JM, Nam HG (1994) Generation of expressed sequence tags of Brassica napus by single-run partial sequencing of random cDNA clones. In: Adams MD, Fields C, Venter JC (eds) Automated DNA sequencing and analysis. Academic Press, London, pp 120–122
Mailer JL, Koontz JW (1981) A study of the induction of cell division in amphibian oocytes by insulin. Dev Biol 85: 309–316
Mochly-Rosen D (1995) Localization of protein kinases by anchoring proteins: a theme in signal transduction. Science 268: 247–251
Mochly-Rosen D, Khaner H, Lopez J (1991) Identification of intracellular receptor proteins for activated protein kinase C. Proc Natl Acad Sci USA 88: 3997–4000
Mochly-Rosen D, Miller KG, Seheller RH, Khaner H, Lopez J, Smith BL (1992) p65 fragments, homologous to the C2 region of protein kinase C, bind to the intracellular receptors for protein kinase C. Biochemistry 31: 8120–8124
Neer EJ, Schmidt CJ, Nambudripad R, Smith TF (1994) The ancient regulatory-protein family of WD-repeat proteins. Nature 371: 297–300
Park YS, Kwak JM, Kwon O-Y, Kim YS, Lee DS, Cho MJ, Lee HH, Nam HG (1993) Generation of expressed sequence tags of random root cDNA clones of Brassica napus by single-run partial sequencing. Plant Physiol 103: 359–370
Park YS, Song O, Kwak JM, Hong SW, Lee HH, Nam HG (1994) Functional complementation of a yeast vesicular transport mutation ypt1-1 by a Brassica napus cDNA clone encoding a small GTP-binding protein. Plant Mol Biol 26: 1725–1735
Pfaff SL, Tamkun MM, Taylor WL (1990) pOEV: a Xenopus oocyte protein expression vector. Anal Biochem 188: 192–199
Rogers SO, Bendish AJ (1985) Extraction of DNA from milligram amounts of fresh herbarium and minifield plant tissues. Plant Mol Biol 5: 69–76
Ron D, Chen C-H, Caldwell J, Jamieson L, Orr E, Mochly-Rosen D (1994) Cloning of an intracellular receptor for protein kinase C: a homolog of the β subunit of G-proteins. Proc Natl Acad Sci USA 91: 839–843
Ron D, Mochly-Rosen D (1994) Agonists and antagonists of protein kinase C function, derived from its binding proteins. J Biol Chem 269: 21395–21398
Ron D, Mochly-Rosen D (1995) An autoregulatory region in protein kinase C: the pseudoanchoring site. Proc Natl Acad Sci USA 92: 492–496
Schloss JA (1990) A Chlamydomonas gene encodes a G protein β subunit-like polypeptide. Mol Gen Genet 221: 443–452
Smith BL, Mochly-Rosen D (1992) Inhibition of protein kinase C function by injection of intracellular receptors for the enzyme. Biochem Biophys Res Comm 188: 1235–1240
Smith LD (1989) The induction of oocyte maturation: transmembrane signaling events and regulation of the cell cycle. Development 107: 685–699
van der Voorn L, Ploegh HL (1992) The WD-40 repeat. FEBS Lett 307: 131–134
Weiss CA, Garnaat CW, Mukai K, Hu Y, Ma H (1994) Isolation of cDNAs encoding guanine nucleotide-binding protein β-subunit homologues from maize (ZGB1) and Arabidopsis (AGB1). Proc Natl Acad Sci USA 91: 9554–9558
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This research was supported in part by a grant from PMBBRC and in part by a grant for promotion of genetic engineering research from the Ministry of Education of Korea. J.M.K. was supported by a fellowship from the Seoam Scholarship Foundation, Seoul, Korea. We thank G. Hoschek for proofreading and G. An for critical reading of this manuscript and for helpful discussion.
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Kwak, J.M., Kim, S.A., Lee, S.K. et al. Insulin-induced maturation of Xenopus oocytes is inhibited by microinjection of a Brassica napus cDNA clone with high similarity to a mammalian receptor for activated protein kinase C. Planta 201, 245–251 (1997). https://doi.org/10.1007/s004250050063
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DOI: https://doi.org/10.1007/s004250050063