Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Jan 1;313(Pt 1):51–55. doi: 10.1042/bj3130051

Annexin II up-regulates cellular levels of p11 protein by a post-translational mechanisms.

A Puisieux 1, J Ji 1, M Ozturk 1
PMCID: PMC1216908  PMID: 8546709

Abstract

Annexin II (p36) and p11, which belong to two different families of calcium-binding proteins, are able to form a heterotetrameric protein complex (p36)2(p11)2 called calpactin I. As these proteins were detectable only in the presence of each other in a variety of cell lines, we studied the mechanisms of regulation of cellular levels of annexin II and p11. In cells expressing p11 messenger RNA, p11 protein is undetectable unless annexin II is also expressed. As an example, the hepatoblastoma HepG2 cell line displays no detectable annexin II nor p11 protein, although it expresses p11 mRNA. The overexpression of annexin II by gene transfer into HepG2 cells leads to the up-regulation of the cellular levels of p11 by a post-translational mechanism. In the presence of annexin II, there is no major change in the p11 transcript levels, but the half-life of the p11 protein is increased more than 6-fold. Thus, the degree of expression of annexin II, which varies according to different states of cellular differentiation and transformation, is an essential factor in the regulation of cellular levels of p11.

Full Text

The Full Text of this article is available as a PDF (333.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baker S. J., Markowitz S., Fearon E. R., Willson J. K., Vogelstein B. Suppression of human colorectal carcinoma cell growth by wild-type p53. Science. 1990 Aug 24;249(4971):912–915. doi: 10.1126/science.2144057. [DOI] [PubMed] [Google Scholar]
  2. Becker T., Weber K., Johnsson N. Protein-protein recognition via short amphiphilic helices; a mutational analysis of the binding site of annexin II for p11. EMBO J. 1990 Dec;9(13):4207–4213. doi: 10.1002/j.1460-2075.1990.tb07868.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blikstad I., Nelson W. J., Moon R. T., Lazarides E. Synthesis and assembly of spectrin during avian erythropoiesis: stoichiometric assembly but unequal synthesis of alpha and beta spectrin. Cell. 1983 Apr;32(4):1081–1091. doi: 10.1016/0092-8674(83)90292-1. [DOI] [PubMed] [Google Scholar]
  4. Bressac B., Galvin K. M., Liang T. J., Isselbacher K. J., Wands J. R., Ozturk M. Abnormal structure and expression of p53 gene in human hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1973–1977. doi: 10.1073/pnas.87.5.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Calabretta B., Venturelli D., Kaczmarek L., Narni F., Talpaz M., Anderson B., Beran M., Baserga R. Altered expression of G1-specific genes in human malignant myeloid cells. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1495–1498. doi: 10.1073/pnas.83.5.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chiang Y., Schneiderman M. H., Vishwanatha J. K. Annexin II expression is regulated during mammalian cell cycle. Cancer Res. 1993 Dec 15;53(24):6017–6021. [PubMed] [Google Scholar]
  7. Creutz C. E. The annexins and exocytosis. Science. 1992 Nov 6;258(5084):924–931. doi: 10.1126/science.1439804. [DOI] [PubMed] [Google Scholar]
  8. Crompton M. R., Moss S. E., Crumpton M. J. Diversity in the lipocortin/calpactin family. Cell. 1988 Oct 7;55(1):1–3. doi: 10.1016/0092-8674(88)90002-5. [DOI] [PubMed] [Google Scholar]
  9. Davidson F. F., Dennis E. A., Powell M., Glenney J. R., Jr Inhibition of phospholipase A2 by "lipocortins" and calpactins. An effect of binding to substrate phospholipids. J Biol Chem. 1987 Feb 5;262(4):1698–1705. [PubMed] [Google Scholar]
  10. Dulis B. H. Regulation of protein expression in differentiation by subunit assembly. Human membrane and secreted IgM. J Biol Chem. 1983 Feb 25;258(4):2181–2187. [PubMed] [Google Scholar]
  11. Emans N., Gorvel J. P., Walter C., Gerke V., Kellner R., Griffiths G., Gruenberg J. Annexin II is a major component of fusogenic endosomal vesicles. J Cell Biol. 1993 Mar;120(6):1357–1369. doi: 10.1083/jcb.120.6.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Erikson E., Shealy D. J., Erikson R. L. Evidence that viral transforming gene products and epidermal growth factor stimulate phosphorylation of the same cellular protein with similar specificity. J Biol Chem. 1981 Nov 25;256(22):11381–11384. [PubMed] [Google Scholar]
  13. Erikson E., Tomasiewicz H. G., Erikson R. L. Biochemical characterization of a 34-kilodalton normal cellular substrate of pp60v-src and an associated 6-kilodalton protein. Mol Cell Biol. 1984 Jan;4(1):77–85. doi: 10.1128/mcb.4.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Frohlich M., Motté P., Galvin K., Takahashi H., Wands J., Ozturk M. Enhanced expression of the protein kinase substrate p36 in human hepatocellular carcinoma. Mol Cell Biol. 1990 Jun;10(6):3216–3223. doi: 10.1128/mcb.10.6.3216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gerke V., Weber K. Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin. EMBO J. 1984 Jan;3(1):227–233. doi: 10.1002/j.1460-2075.1984.tb01789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gerke V., Weber K. The regulatory chain in the p36-kd substrate complex of viral tyrosine-specific protein kinases is related in sequence to the S-100 protein of glial cells. EMBO J. 1985 Nov;4(11):2917–2920. doi: 10.1002/j.1460-2075.1985.tb04023.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Glenney J. R., Jr Calpactins: calcium-regulated membrane-skeletal proteins. Bioessays. 1987 Oct;7(4):173–175. doi: 10.1002/bies.950070408. [DOI] [PubMed] [Google Scholar]
  18. Glenney J. R., Jr, Tack B. F. Amino-terminal sequence of p36 and associated p10: identification of the site of tyrosine phosphorylation and homology with S-100. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7884–7888. doi: 10.1073/pnas.82.23.7884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Glenney J. R., Jr, Tack B., Powell M. A. Calpactins: two distinct Ca++-regulated phospholipid- and actin-binding proteins isolated from lung and placenta. J Cell Biol. 1987 Mar;104(3):503–511. doi: 10.1083/jcb.104.3.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gould K. L., Cooper J. A., Hunter T. The 46,000-dalton tyrosine protein kinase substrate is widespread, whereas the 36,000-dalton substrate is only expressed at high levels in certain rodent tissues. J Cell Biol. 1984 Feb;98(2):487–497. doi: 10.1083/jcb.98.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harder T., Gerke V. The subcellular distribution of early endosomes is affected by the annexin II2p11(2) complex. J Cell Biol. 1993 Dec;123(5):1119–1132. doi: 10.1083/jcb.123.5.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Harder T., Kube E., Gerke V. Cloning and characterization of the human gene encoding p11: structural similarity to other members of the S-100 gene family. Gene. 1992 Apr 15;113(2):269–274. doi: 10.1016/0378-1119(92)90406-f. [DOI] [PubMed] [Google Scholar]
  23. Hunter T. The Ca2+/phospholipid-binding proteins of the submembraneous skeleton. Adv Exp Med Biol. 1988;234:169–193. doi: 10.1007/978-1-4757-1980-2_12. [DOI] [PubMed] [Google Scholar]
  24. Isacke C. M., Trowbridge I. S., Hunter T. Modulation of p36 phosphorylation in human cells: studies using anti-p36 monoclonal antibodies. Mol Cell Biol. 1986 Jul;6(7):2745–2751. doi: 10.1128/mcb.6.7.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jindal H. K., Chaney W. G., Anderson C. W., Davis R. G., Vishwanatha J. K. The protein-tyrosine kinase substrate, calpactin I heavy chain (p36), is part of the primer recognition protein complex that interacts with DNA polymerase alpha. J Biol Chem. 1991 Mar 15;266(8):5169–5176. [PubMed] [Google Scholar]
  26. Johnsson N., Marriott G., Weber K. p36, the major cytoplasmic substrate of src tyrosine protein kinase, binds to its p11 regulatory subunit via a short amino-terminal amphiphatic helix. EMBO J. 1988 Aug;7(8):2435–2442. doi: 10.1002/j.1460-2075.1988.tb03089.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Johnsson N., Nguyen Van P., Söling H. D., Weber K. Functionally distinct serine phosphorylation sites of p36, the cellular substrate of retroviral protein kinase; differential inhibition of reassociation with p11. EMBO J. 1986 Dec 20;5(13):3455–3460. doi: 10.1002/j.1460-2075.1986.tb04669.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Klee C. B. Ca2+-dependent phospholipid- (and membrane-) binding proteins. Biochemistry. 1988 Sep 6;27(18):6645–6653. doi: 10.1021/bi00418a001. [DOI] [PubMed] [Google Scholar]
  29. Kligman D., Hilt D. C. The S100 protein family. Trends Biochem Sci. 1988 Nov;13(11):437–443. doi: 10.1016/0968-0004(88)90218-6. [DOI] [PubMed] [Google Scholar]
  30. Merlie J. P., Lindstrom J. Assembly in vivo of mouse muscle acetylcholine receptor: identification of an alpha subunit species that may be an assembly intermediate. Cell. 1983 Oct;34(3):747–757. doi: 10.1016/0092-8674(83)90531-7. [DOI] [PubMed] [Google Scholar]
  31. Merlie J. P., Sebbane R., Tzartos S., Lindstrom J. Inhibition of glycosylation with tunicamycin blocks assembly of newly synthesized acetylcholine receptor subunits in muscle cells. J Biol Chem. 1982 Mar 10;257(5):2694–2701. [PubMed] [Google Scholar]
  32. Ozaki T., Sakiyama S. Molecular cloning of rat calpactin I heavy-chain cDNA whose expression is induced in v-src-transformed rat culture cell lines. Oncogene. 1993 Jun;8(6):1707–1710. [PubMed] [Google Scholar]
  33. Powell M. A., Glenney J. R. Regulation of calpactin I phospholipid binding by calpactin I light-chain binding and phosphorylation by p60v-src. Biochem J. 1987 Oct 15;247(2):321–328. doi: 10.1042/bj2470321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Puisieux A., Ponchel F., Ozturk M. p53 as a growth suppressor gene in HBV-related hepatocellular carcinoma cells. Oncogene. 1993 Feb;8(2):487–490. [PubMed] [Google Scholar]
  35. Radke K., Gilmore T., Martin G. S. Transformation by Rous sarcoma virus: a cellular substrate for transformation-specific protein phosphorylation contains phosphotyrosine. Cell. 1980 Oct;21(3):821–828. doi: 10.1016/0092-8674(80)90445-6. [DOI] [PubMed] [Google Scholar]
  36. Schlaepfer D. D., Haigler H. T. Expression of annexins as a function of cellular growth state. J Cell Biol. 1990 Jul;111(1):229–238. doi: 10.1083/jcb.111.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Spano F., Raugei G., Palla E., Colella C., Melli M. Characterization of the human lipocortin-2-encoding multigene family: its structure suggests the existence of a short amino acid unit undergoing duplication. Gene. 1990 Nov 15;95(2):243–251. doi: 10.1016/0378-1119(90)90367-z. [DOI] [PubMed] [Google Scholar]
  38. Stuve L. L., Myers R. M. A directly repeated sequence in the beta-globin promoter regulates transcription in murine erythroleukemia cells. Mol Cell Biol. 1990 Mar;10(3):972–981. doi: 10.1128/mcb.10.3.972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vishwanatha J. K., Kumble S. Involvement of annexin II in DNA replication: evidence from cell-free extracts of Xenopus eggs. J Cell Sci. 1993 Jun;105(Pt 2):533–540. doi: 10.1242/jcs.105.2.533. [DOI] [PubMed] [Google Scholar]
  40. Wallner B. P., Mattaliano R. J., Hession C., Cate R. L., Tizard R., Sinclair L. K., Foeller C., Chow E. P., Browing J. L., Ramachandran K. L. Cloning and expression of human lipocortin, a phospholipase A2 inhibitor with potential anti-inflammatory activity. Nature. 1986 Mar 6;320(6057):77–81. doi: 10.1038/320077a0. [DOI] [PubMed] [Google Scholar]
  41. Zokas L., Glenney J. R., Jr The calpactin light chain is tightly linked to the cytoskeletal form of calpactin I: studies using monoclonal antibodies to calpactin subunits. J Cell Biol. 1987 Nov;105(5):2111–2121. doi: 10.1083/jcb.105.5.2111. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES