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PMID: 10202149 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and characterization of a cDNA encoding an A-kinase anchoring protein located in the centrosome, AKAP450.

The EMBO journal ·Vol. 18 ·No. 7 ·1999-04-01 ·Pages 1858-68

Witczak O, Skålhegg BS, Keryer G, Bornens M, Taskén K, Jahnsen T, Orstavik S

Abstract

A combination of protein kinase A type II (RII) overlay screening, database searches and PCR was used to identify a centrosomal A-kinase anchoring protein. A cDNA with an 11.7 kb open reading frame was characterized and found to correspond to 50 exons of genomic sequence on human chromosome 7q21-22. This cDNA clone encoded a 3908 amino acid protein of 453 kDa, that was designated AKAP450 (DDBJ/EMBL/GenBank accession No. AJ131693). Sequence comparison demonstrated that the open reading frame contained a previously characterized cDNA encoding Yotiao, as well as the human homologue of AKAP120. Numerous coiled-coil structures were predicted from AKAP450, and weak homology to pericentrin, giantin and other structural proteins was observed. A putative RII-binding site was identified involving amino acid 2556 of AKAP450 by mutation analysis combined with RII overlay and an amphipatic helix was predicted in this region. Immunoprecipitation of RII from RIPA-buffer extracts of HeLa cells demonstrated co-precipitation of AKAP450. By immunofluorecent labeling with specific antibodies it was demonstrated that AKAP450 localized to centrosomes. Furthermore, AKAP450 was shown to co-purify in centrosomal preparations. The observation of two mRNAs and several splice products suggests additional functions for the AKAP450 gene.

MeSH Terms
A Kinase Anchor Proteins Adaptor Proteins, Signal Transducing Amino Acid Sequence Base Sequence Binding Sites Carrier Proteins Cell Line Centrosome/metabolism Cloning, Molecular Cyclic AMP-Dependent Protein Kinase Type II Cyclic AMP-Dependent Protein Kinases/metabolism Cytoskeletal Proteins DNA Primers/genetics DNA, Complementary/genetics Exons Female Humans Introns Microtubule-Associated Proteins/genetics,isolation & purification,metabolism Molecular Sequence Data Pregnancy RNA, Messenger/genetics,metabolism Subcellular Fractions/metabolism Tissue Distribution
Chemicals
A Kinase Anchor Proteins AKAP9 protein, human Adaptor Proteins, Signal Transducing Carrier Proteins Cytoskeletal Proteins DNA Primers DNA, Complementary Microtubule-Associated Proteins RNA, Messenger Cyclic AMP-Dependent Protein Kinase Type II Cyclic AMP-Dependent Protein Kinases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Witczak O
Institute of Medical Biochemistry, Faculty of Medicine, University of Oslo, PO Box 1112, Blindern N-0317 Oslo, Norway.
Skålhegg B S
Keryer G
Bornens M
Taskén K
Jahnsen T
Orstavik S
References (38)
38 references, click to expand
  1. Cyclic nucleotide-dependent protein kinases.
    Pharmacol Ther. 1991;50(1):123-45 PMID: 1653962
  2. Interaction of the regulatory subunit (RII) of cAMP-dependent protein kinase with RII-anchoring proteins occurs through an amphipathic helix binding motif.
    J Biol Chem. 1991 Aug 5;266(22):14188-92 PMID: 1860836
  3. A high-affinity binding protein for the regulatory subunit of cAMP-dependent protein kinase II in the centrosome of human cells.
    Exp Cell Res. 1993 Feb;204(2):230-40 PMID: 8440320
  4. Novel isozymes of cAMP-dependent protein kinase exist in human cells due to formation of RI alpha-RI beta heterodimeric complexes.
    J Biol Chem. 1993 Oct 5;268(28):21276-83 PMID: 8407966
  5. Location of cAMP-dependent protein kinase type I with the TCR-CD3 complex.
    Science. 1994 Jan 7;263(5143):84-7 PMID: 8272870
  6. Pericentrin, a highly conserved centrosome protein involved in microtubule organization.
    Cell. 1994 Feb 25;76(4):639-50 PMID: 8124707
  7. Structure and function of cyclic nucleotide-dependent protein kinases.
    Annu Rev Physiol. 1994;56:237-72 PMID: 8010741
  8. RNA localization along the anteroposterior axis of the Drosophila oocyte requires PKA-mediated signal transduction to direct normal microtubule organization.
    Genes Dev. 1994 Dec 15;8(24):2986-95 PMID: 7528157
  9. A kinase anchor proteins and the intracellular targeting of signals carried by cyclic AMP.
    Biochim Biophys Acta. 1994 Dec 30;1224(3):467-79 PMID: 7803506
  10. Molecular characterisation of ninein, a new coiled-coil protein of the centrosome.
    J Cell Sci. 1996 Jan;109 ( Pt 1):179-90 PMID: 8834802
  11. Properties of A-kinase anchoring proteins.
    Biochem Soc Trans. 1996 Nov;24(4):986-91 PMID: 8968497
  12. Identification and characterization of a novel A-kinase-anchoring protein (AKAP120) from rabbit gastric parietal cells.
    Biochem J. 1997 Mar 15;322 ( Pt 3):801-8 PMID: 9148752
  13. The human mitochondrial transcription termination factor (mTERF) is a multizipper protein but binds to DNA as a monomer, with evidence pointing to intramolecular leucine zipper interactions.
    EMBO J. 1997 Mar 3;16(5):1066-79 PMID: 9118945
  14. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.
    Nucleic Acids Res. 1997 Sep 1;25(17):3389-402 PMID: 9254694
  15. Regulation of microtubule dynamics by extracellular signals: cAMP-dependent protein kinase switches off the activity of oncoprotein 18 in intact cells.
    J Cell Biol. 1998 Jan 12;140(1):131-41 PMID: 9425161
  16. A centrosomal function for the human Nek2 protein kinase, a member of the NIMA family of cell cycle regulators.
    EMBO J. 1998 Jan 15;17(2):470-81 PMID: 9430639
  17. Molecular cloning, chromosomal localization, and cell cycle-dependent subcellular distribution of the A-kinase anchoring protein, AKAP95.
    Exp Cell Res. 1998 Feb 1;238(2):305-16 PMID: 9473338
  18. Yotiao, a novel protein of neuromuscular junction and brain that interacts with specific splice variants of NMDA receptor subunit NR1.
    J Neurosci. 1998 Mar 15;18(6):2017-27 PMID: 9482789
  19. Implications for cAMP-dependent protein kinase in the maintenance of the interphase state.
    Prog Cell Cycle Res. 1995;1:241-53 PMID: 9552367
  20. A homologue of Drosophila aurora kinase is oncogenic and amplified in human colorectal cancers.
    EMBO J. 1998 Jun 1;17(11):3052-65 PMID: 9606188
  21. C-Nap1, a novel centrosomal coiled-coil protein and candidate substrate of the cell cycle-regulated protein kinase Nek2.
    J Cell Biol. 1998 Jun 29;141(7):1563-74 PMID: 9647649
  22. Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation.
    Nat Genet. 1998 Oct;20(2):189-93 PMID: 9771714
  23. The stathmin phosphoprotein family: intracellular localization and effects on the microtubule network.
    J Cell Sci. 1998 Nov;111 ( Pt 22):3333-46 PMID: 9788875
  24. Mitosis-specific phosphorylation and subcellular redistribution of the RIIalpha regulatory subunit of cAMP-dependent protein kinase.
    J Biol Chem. 1998 Dec 18;273(51):34594-602 PMID: 9852131
  25. Human T cell hybridomas secreting factors for IgA-specific help, polyclonal B cell activation, and B cell proliferation.
    J Exp Med. 1982 Dec 1;156(6):1860-5 PMID: 6983562
  26. High-affinity binding of the regulatory subunit (RII) of cAMP-dependent protein kinase to microtubule-associated and other cellular proteins.
    Proc Natl Acad Sci U S A. 1984 Nov;81(21):6723-7 PMID: 6093118
  27. Identification and differential expression of two forms of regulatory subunits (RII) of cAMP-dependent protein kinase II in Friend erythroleukemic cells. Differentiation and 8-bromo-cAMP elicit a large and selective increase in the rate of biosynthesis of only one type of RII.
    J Biol Chem. 1985 May 25;260(10):6296-303 PMID: 2581952
  28. Cyclic-AMP-dependent protein kinase type II is associated with the Golgi complex and with centrosomes.
    Cell. 1985 Jul;41(3):1039-51 PMID: 2988780
  29. Microtubule-associated protein 1B: identification of a major component of the neuronal cytoskeleton.
    Proc Natl Acad Sci U S A. 1985 Aug;82(16):5404-8 PMID: 3895231
  30. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes.
    Cell. 1986 Jan 31;44(2):283-92 PMID: 3943125
  31. Heterogeneous distribution of the cAMP receptor protein RII in the nervous system: evidence for its intracellular accumulation on microtubules, microtubule-organizing centers, and in the area of the Golgi complex.
    J Cell Biol. 1986 Jul;103(1):189-203 PMID: 3522603
  32. Identification of centrosomal proteins in a human lymphoblastic cell line.
    EMBO J. 1986 Oct;5(10):2545-50 PMID: 3536479
  33. Structural and chemical characterization of isolated centrosomes.
    Cell Motil Cytoskeleton. 1987;8(3):238-49 PMID: 3690689
  34. High affinity binding protein for the regulatory subunit of cAMP-dependent protein kinase II-B. Cloning, characterization, and expression of cDNAs for rat brain P150.
    J Biol Chem. 1989 Mar 15;264(8):4648-56 PMID: 2538452
  35. p34cdc2 is located in both nucleus and cytoplasm; part is centrosomally associated at G2/M and enters vesicles at anaphase.
    EMBO J. 1989 Dec 20;8(13):3985-95 PMID: 2686978
  36. In vitro effects on microtubule dynamics of purified Xenopus M phase-activated MAP kinase.
    Nature. 1991 Jan 17;349(6306):251-4 PMID: 1702878
  37. Predicting coiled coils from protein sequences.
    Science. 1991 May 24;252(5009):1162-4 PMID: 2031185
  38. Identification, purification, and characterization of subunits of cAMP-dependent protein kinase in human testis. Reverse mobilities of human RII alpha and RII beta on sodium dodecyl sulfate-polyacrylamide gel electrophoresis compared with rat and bovine RIIs.
    J Biol Chem. 1992 Mar 15;267(8):5374-9 PMID: 1544918
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/18.7.1858
Published
1999-04-01
Pages
1858-68
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171271
Subset
IM
Databases
GENBANK
AJ131693
External Links
PubMed source DOI: 10.1093/emboj/18.7.1858 PMC Full Text View journal details
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