Abstract
Phosphoprotein phosphatases are central regulatory components of the cell cycle in eukaryotes. We report the cloning and sequencing of an alfalfa phosphoprotein phosphatase type 2A (pp2aMs) cDNA. The predicted protein sequence shows high similarity to PP2A from Brassica napus, rabbit and Drosophila. No changes in pp2aMs mRNA abundance during the cell cycle were found. During growth of a batch cell culture, mRNA levels decreased gradually. In planta, all organs contained pp2a transcripts but maximal mRNA levels were detected in stems. Since Southern analysis indicated the presence of a small pp2a gene family in alfalfa, it appears that different subtypes may have specialized roles in various tissues and developmental situations which await characterization.
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Arino J, Woon CW, Brautigan DL, Miller JR, TB, Johnson GL (1988) Human liver phosphatase 2A:cDNA and amino acid sequence of two catalytic subunit isotypes. Proc Natl Acad Sci USA 85:4252–4256
Axton JM, Dombradi V, Cohen PTW, Glover D (1990) One of the protein phosphatase 1 isoenzymes in Drosophila is essential for mitosis. Cell 63:33–46
Booher R, Beach D (1987) Interaction between cdc13 + and cdc2 + in the control of mitosis in fission yeast; dissociation of the G1 and G2 roles of the cdc2 + protein kinase. EMBO J 6:3441–3447
Booher R, Beach D (1988) Involvement of cdc13 + in mitotic control in Schizosaccharomyces pombe: possible interaction of the gene product with microtubules. EMBO J 7:2321–2327
Chomczynski P, Sacchi N (1986) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Cohen P, Holmes CFB, Tsukitani Y (1990) Okadaic acid: a new probe for the study of cellular regulation. Trends Biochem 15:98–102
Doonan JH (1991) Cycling plant cells. Plant J 1: 129–132
Doonan JH, Morris R (1989) The bimG gene of Aspergillus nidulans, required for completion of anaphase, encodes a homolog of mammalian phosphoprotein phosphatase 1. Cell 57:987–996
Dunphy WG, Newport JW (1989) Fission yeast p13 blocks mitotic activation and tyrosine dephosphorylation of the Xenopus cdc2 protein kinase. Cell 58:181–191
Felix MA, Cohen P, Karsenti E (1990) Cdc2 H1 kinase is negatively regulated by a type 2A phosphatase in the Xenopus early embryonic cell cycle: evidence from the effects of okadaic acid. EMBO J 9:675–683
Gautier J, Matsukawa T, Nurse P, Maller J (1989) Dephosphorylation and activation of Xenopus p34 protein kinase during the cell cycle. Nature 339:626–629
Gould SJ, Subramani S, Scheffler IE (1989) Use of the DNA polymerase chain reaction for homology probing: Isolation of partial cDNA or genomic clones encoding the iron-sulfur protein of succinate dehydrogenase from several species. Proc Natl Acad Sci USA 86:1934–1938
Györgyey J, Gartner A, Nemeth K, Magyar Z, Hirt H, Heberle-Bors E, Dudits D (1991) Alfalfa heat shock genes are differentially expressed during somatic embryogenesis. Plant Mol Biol 16:999–1007
Healy AM, Zolnierowicz S, Stapleton AE, Goebl M, DePaoli-Roach AA, Pringle JR (1991) CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: Identification, characterization, and homology to the B subunit of mammalian type 2A protein phosphatase. Mol Cell Biol 11:5767–5780
Kinoshita N, Ohkura H, Yanagida M (1990) Distinct, essential roles of type 1 and 2A protein phosphatases in the control of the fission yeast cell division cycle. Cell 63:405–415
MacKintosh C, Cohen P (1989) Identification of high levels of type 1 and type 2A protein phosphatases in higher plants. Biochem J 262:335–339
MacKintosh C, Coggins J, Cohen P (1991) Plant protein phosphatases. Biochem J 273:733–738
MacKintosh RW, Haycox G, Hardie DG, Cohen P (1990) Identification by molecular cloning of two cDNA sequences from the plant Brassica napus which are very similar to mammalian protein phosphatases-1 and -2A. FEBS Lett 276:156–160
Minshull J, Blow JJ, Hunt T (1989) Translation of cyclin mRNA is necessary for extracts of activated Xenopus eggs to enter mitosis. Cell 56:947–956
Mumby MC, Walter G (1991) Protein phosphatases and DNA tumor viruses: Transformation through the back door? Cell Regul 2:589–598
Murashige T, Skoog F (1962) A revised medium for rapid growth of and bioassays with tissue cultures. Physiol Plant 15:473–479
Murray AM, Kirschner MW (1989) Cyclin synthesis drives the early embryonic cell cycle. Nature 339:275–280
Murray AM, Solomon MJ, Kirschner MW (1989) The role of cyclin synthesis and degradation in the control of maturation promoter activity. Nature 339:280–286
Nurse P (1990) Universal control mechanism regulating onset of M-phase. Nature 344:503–507
Ohkura H, Kinoshita N, Miyatani S, Toda T, Yanagida M (1989) The fission yeast dis2 gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases. Cell 57:997–1007
Páy A, Heberle-Bors E, Hirt H (1992) An alfalfa cDNA encodes a protein with homology to translationally controlled human tumor protein. Plant Mol Biol 19:501–503
Pfosser M (1989) Improved method for critical comparison of cell cycle data of asynchronously dividing and synchronized cell cultures of Nicotiana tabacum. J Plant Physiol 134:741–745
Ronne H, Carlberg M, Hu G-Z, Nehlin JO (1991) Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Mol Cell Biol 11:4876–4884
Ruediger R, Van Wart Hood JE, Mumby M, Walter G (1991) Constant expression and activity of protein phosphatase 2A in synchronized cells. Mol Cell Biol 11:4282–4285
Solomon MJ, Glotzer M, Lee TH, Philippe M, Kirschner MW (1990) Cyclin activation of p34cdc2. Cell 63:1013–1024
Smythe C, Newport JW (1992) Coupling of mitosis to the completion of S phase in Xenopus occurs via modulation of the tyrosine kinase that phosphorylates p34cdc2. Cell 68:787–797
Sutton A, Immanuel D, Arndt KT (1991) The SIT4 protein phosphatase functions in late G1 for progression into S phase. Mol Cell Biol 11:2133–2148
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Communicated by A. Kondorosi
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Pirck, M., Páy, A., Heberle-Bors, E. et al. Isolation and characterization of a phosphoprotein phosphatase type 2A gene from alfalfa. Molec. Gen. Genet. 240, 126–131 (1993). https://doi.org/10.1007/BF00276891
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DOI: https://doi.org/10.1007/BF00276891