Molecular Cloning of the Genes Encoding the PR55/Bβ/δ Regulatory Subunits for PP-2A and Analysis of Their Functions in Regulating Development of Goldfish, Carassius Auratus

Overview

Journal Gene Regul Syst Bio

Publisher Sage Publications

Date 2011 Jan 20

PMID 21245947

Citations 2

Authors

Jun-Qiong Zhao

Si-Si Xie

Wen-Bin Liu

Ya-Mei Xiao

Xiao-Ming Zeng

Mi Deng

Lili Gong

Jin-Ping Liu

Pei-Chao Chen

Jie Zhou

Xiao-Hui Hu

Jia-Han Lv

Xiang-Qian Yu

Dao Wang

Chi Li

Yun-Lei Peng

Gao-Peng Liao

Yun Liu

David Wan-Cheng Li

Affiliations

Soon will be listed here.

Abstract

The protein phosphatase-2A (PP-2A), one of the major phosphatases in eukaryotes, is a heterotrimer, consisting of a scaffold A subunit, a catalytic C subunit and a regulatory B subunit. Previous studies have shown that besides regulating specific PP-2A activity, various B subunits encoded by more than 16 different genes, may have other functions. To explore the possible roles of the regulatory subunits of PP-2A in vertebrate development, we have cloned the PR55/B family regulatory subunits: β and δ, analyzed their tissue specific and developmental expression patterns in Goldfish ( Carassius auratus). Our results revealed that the full-length cDNA for PR55/Bβ consists of 1940 bp with an open reading frame of 1332 nucleotides coding for a deduced protein of 443 amino acids. The full length PR55/Bδ cDNA is 2163 bp containing an open reading frame of 1347 nucleotides encoding a deduced protein of 448 amino acids. The two isoforms of PR55/B display high levels of sequence identity with their counterparts in other species. The PR55/Bβ mRNA and protein are detected in brain and heart. In contrast, the PR55/Bδ is expressed in all 9 tissues examined at both mRNA and protein levels. During development of goldfish, the mRNAs for PR55/Bβ and PR55/Bδ show distinct patterns. At the protein level, PR55/Bδ is expressed at all developmental stages examined, suggesting its important role in regulating goldfish development. Expression of the PR55/Bδ anti-sense RNA leads to significant downregulation of PR55/Bδ proteins and caused severe abnormality in goldfish trunk and eye development. Together, our results suggested that PR55/Bδ plays an important role in governing normal trunk and eye formation during goldfish development.

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References

Uemura T, Shiomi K, Togashi S, Takeichi M . Mutation of twins encoding a regulator of protein phosphatase 2A leads to pattern duplication in Drosophila imaginal discs. Genes Dev. 1993; 7(3):429-40. DOI: 10.1101/gad.7.3.429. View

Hunter T . Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell. 1995; 80(2):225-36. DOI: 10.1016/0092-8674(95)90405-0. View

Ecker J, Davis R . Inhibition of gene expression in plant cells by expression of antisense RNA. Proc Natl Acad Sci U S A. 1986; 83(15):5372-6. PMC: 386288. DOI: 10.1073/pnas.83.15.5372. View

McGarry T, Lindquist S . Inhibition of heat shock protein synthesis by heat-inducible antisense RNA. Proc Natl Acad Sci U S A. 1986; 83(2):399-403. PMC: 322866. DOI: 10.1073/pnas.83.2.399. View

Li X, Virshup D . Two conserved domains in regulatory B subunits mediate binding to the A subunit of protein phosphatase 2A. Eur J Biochem. 2002; 269(2):546-52. DOI: 10.1046/j.0014-2956.2001.02680.x. View

Millward T, Zolnierowicz S, Hemmings B . Regulation of protein kinase cascades by protein phosphatase 2A. Trends Biochem Sci. 1999; 24(5):186-91. DOI: 10.1016/s0968-0004(99)01375-4. View

Chen H, Han W, Deng M, Qin J, Yuan D, Liu J . Transcriptional regulation of PP2A-A alpha is mediated by multiple factors including AP-2alpha, CREB, ETS-1, and SP-1. PLoS One. 2009; 4(9):e7019. PMC: 2736573. DOI: 10.1371/journal.pone.0007019. View

Qin J, Chen H, Yan Q, Deng M, Liu J, Doerge S . Protein phosphatase-2A is a target of epigallocatechin-3-gallate and modulates p53-Bak apoptotic pathway. Cancer Res. 2008; 68(11):4150-62. DOI: 10.1158/0008-5472.CAN-08-0839. View

Suzuki M, Imanishi S, Dohmae N, Asanuma M, Matsumoto S . Identification of a male-specific RNA binding protein that regulates sex-specific splicing of Bmdsx by increasing RNA binding activity of BmPSI. Mol Cell Biol. 2010; 30(24):5776-86. PMC: 3004276. DOI: 10.1128/MCB.00444-10. View

10.

Mayer-Jaekel R, Ohkura H, Gomes R, Sunkel C, Baumgartner S, Hemmings B . The 55 kd regulatory subunit of Drosophila protein phosphatase 2A is required for anaphase. Cell. 1993; 72(4):621-33. DOI: 10.1016/0092-8674(93)90080-a. View

11.

Xiao Y, Chen L, Liu J, Liu W, Chen H, Zou L . Contrast expression patterns of JNK1 during sex reversal of the rice-field eel. J Exp Zool B Mol Dev Evol. 2009; 314(3):242-56. DOI: 10.1002/jez.b.21332. View

12.

Hatano Y, Shima H, Haneji T, Miura A, Sugimura T, Nagao M . Expression of PP2A B regulatory subunit beta isotype in rat testis. FEBS Lett. 1993; 324(1):71-5. DOI: 10.1016/0014-5793(93)81535-8. View

13.

Moorhead G, Trinkle-Mulcahy L, Ulke-Lemee A . Emerging roles of nuclear protein phosphatases. Nat Rev Mol Cell Biol. 2007; 8(3):234-44. DOI: 10.1038/nrm2126. View

14.

Goedert M, Cohen E, Jakes R, Cohen P . p42 MAP kinase phosphorylation sites in microtubule-associated protein tau are dephosphorylated by protein phosphatase 2A1. Implications for Alzheimer's disease [corrected]. FEBS Lett. 1992; 312(1):95-9. DOI: 10.1016/0014-5793(92)81418-l. View

15.

Strack S, Chang D, Zaucha J, Colbran R, Wadzinski B . Cloning and characterization of B delta, a novel regulatory subunit of protein phosphatase 2A. FEBS Lett. 1999; 460(3):462-6. DOI: 10.1016/s0014-5793(99)01377-0. View

16.

Fu H, Ma H, Zheng C, Lu J, Yu X, Li C . Molecular cloning and differential expression patterns of the regulatory subunit B' gene of PP2A in goldfish, Carassius auratus. Sci China C Life Sci. 2009; 52(8):724-32. DOI: 10.1007/s11427-009-0094-4. View

17.

Li D, Liu J, Mao Y, Xiang H, Wang J, Ma W . Calcium-activated RAF/MEK/ERK signaling pathway mediates p53-dependent apoptosis and is abrogated by alpha B-crystallin through inhibition of RAS activation. Mol Biol Cell. 2005; 16(9):4437-53. PMC: 1196350. DOI: 10.1091/mbc.e05-01-0010. View

18.

Yamashiro S, Yamakita Y, Ishikawa R, Matsumura F . Mitosis-specific phosphorylation causes 83K non-muscle caldesmon to dissociate from microfilaments. Nature. 1990; 344(6267):675-8. DOI: 10.1038/344675a0. View

19.

Xing Y, Xu Y, Chen Y, Jeffrey P, Chao Y, Lin Z . Structure of protein phosphatase 2A core enzyme bound to tumor-inducing toxins. Cell. 2006; 127(2):341-53. DOI: 10.1016/j.cell.2006.09.025. View

20.

Ferrigno P, LANGAN T, Cohen P . Protein phosphatase 2A1 is the major enzyme in vertebrate cell extracts that dephosphorylates several physiological substrates for cyclin-dependent protein kinases. Mol Biol Cell. 1993; 4(7):669-77. PMC: 300977. DOI: 10.1091/mbc.4.7.669. View