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PMID: 9372968 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

I kappaB alpha physically interacts with a cytoskeleton-associated protein through its signal response domain.

Molecular and cellular biology ·Vol. 17 ·No. 12 ·1997-12-00 ·Pages 7375-85

Crépieux P, Kwon H, Leclerc N, Spencer W, Richard S, Lin R, Hiscott J

Abstract

The I kappaB alpha protein is a key molecular target involved in the control of NF-kappaB/Rel transcription factors during viral infection or inflammatory reactions. This NF-kappaB-inhibitory factor is regulated by posttranslational phosphorylation and ubiquitination of its amino-terminal signal response domain that targets I kappaB alpha for rapid proteolysis by the 26S proteasome. In an attempt to identify regulators of the I kappaB alpha inhibitory activity, we undertook a yeast two-hybrid genetic screen, using the amino-terminal end of I kappaB alpha as bait, and identified 12 independent interacting clones. Sequence analysis identified some of these cDNA clones as Dlc-1, a sequence encoding a small, 9-kDa human homolog of the outer-arm dynein light-chain protein. In the two-hybrid assay, Dlc-1 also interacted with full-length I kappaB alpha protein but not with N-terminal-deletion-containing versions of I kappaB alpha. I kappaB alpha interacted in vitro with a glutathione S-transferase-Dlc-1 fusion protein, and RelA(p65) did not displace this association, demonstrating that p65 and Dlc-1 contact different protein motifs of I kappaB alpha. Importantly, in HeLa and 293 cells, endogenous and transfected I kappaB alpha coimmunoprecipitated with Myc-tagged or endogenous Dlc-1. Indirect immunofluorescence analyzed by confocal microscopy indicated that Dlc-1 and I kappaB alpha colocalized with both nuclear and cytoplasmic distribution. Furthermore, Dlc-1 and I kappaB alpha were found to associate with the microtubule organizing center, a perinuclear region from which microtubules radiate. Likewise, I kappaB alpha colocalized with alpha-tubulin filaments. Taken together, these results highlight an intriguing interaction between the I kappaB alpha protein and the human homolog of a member of the dynein family of motor proteins and provide a potential link between cytoskeleton dynamics and gene regulation.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites/genetics Cell Line Cell Nucleus/metabolism Cytoplasm/metabolism Cytoskeletal Proteins/chemistry,genetics,metabolism DNA Primers/genetics DNA-Binding Proteins/chemistry,genetics,metabolism Dyneins/chemistry,genetics,metabolism Fluorescent Antibody Technique, Indirect Humans I-kappa B Proteins Microscopy, Confocal Microtubules/metabolism Molecular Sequence Data NF-KappaB Inhibitor alpha Polymerase Chain Reaction Protein Sorting Signals/chemistry,genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism Sequence Homology, Amino Acid Transfection
Chemicals
Cytoskeletal Proteins DNA Primers DNA-Binding Proteins I-kappa B Proteins NFKBIA protein, human Protein Sorting Signals NF-KappaB Inhibitor alpha Dyneins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Crépieux P
Terry Fox Molecular Oncology Group, Lady Davis Institute for Medical Research, Department of Medicine, McGill University, Montreal, Que., Canada.
Kwon H
Leclerc N
Spencer W
Richard S
Lin R
Hiscott J
References (69)
69 references, click to expand
  1. Cytoplasmic dynein (ddlc1) mutations cause morphogenetic defects and apoptotic cell death in Drosophila melanogaster.
    Mol Cell Biol. 1996 May;16(5):1966-77 PMID: 8628263
  2. The role of the C-terminal domain of I kappa B alpha in protein degradation and stabilization.
    J Biol Chem. 1996 May 3;271(18):10690-6 PMID: 8631876
  3. Cell cycle regulation of dynein association with membranes modulates microtubule-based organelle transport.
    J Cell Biol. 1996 May;133(3):585-93 PMID: 8636233
  4. Phosphorylation of IkappaBalpha in the C-terminal PEST domain by casein kinase II affects intrinsic protein stability.
    Mol Cell Biol. 1996 Apr;16(4):1401-9 PMID: 8657113
  5. Identification of lysine residues required for signal-induced ubiquitination and degradation of I kappa B-alpha in vivo.
    Oncogene. 1996 Jun 6;12(11):2425-35 PMID: 8649784
  6. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier.
    Curr Genet. 1989 Dec;16(5-6):339-46 PMID: 2692852
  7. Analysis of cDNA for human erythrocyte ankyrin indicates a repeated structure with homology to tissue-differentiation and cell-cycle control proteins.
    Nature. 1990 Mar 1;344(6261):36-42 PMID: 2137557
  8. A sequestered pool of aminoacyl-tRNA in mammalian cells.
    Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3601-4 PMID: 1565655
  9. Direct association of pp40/I kappa B beta with rel/NF-kappa B transcription factors: role of ankyrin repeats in the inhibition of DNA binding activity.
    Proc Natl Acad Sci U S A. 1992 May 15;89(10):4333-7 PMID: 1533932
  10. The C terminus of the NF-kappa B p50 precursor and an I kappa B isoform contain transcription activation domains.
    Nucleic Acids Res. 1992 May 25;20(10):2453-8 PMID: 1598203
  11. I kappa B interacts with the nuclear localization sequences of the subunits of NF-kappa B: a mechanism for cytoplasmic retention.
    Genes Dev. 1992 Oct;6(10):1899-913 PMID: 1340770
  12. Tyrosine-phosphorylation of tubulin during monocytic differentiation of HL-60 cells.
    J Immunol. 1993 Jan 15;150(2):585-93 PMID: 7678277
  13. Immunofluorescent localization of the ubiquitin-activating enzyme, E1, to the nucleus and cytoskeleton.
    Am J Physiol. 1993 Jan;264(1 Pt 1):C93-102 PMID: 8430776
  14. Actions of Rho family small G proteins and p21-activated protein kinases on mitogen-activated protein kinase family members.
    Mol Cell Biol. 1996 Jul;16(7):3707-13 PMID: 8668187
  15. Brain cytoplasmic and flagellar outer arm dyneins share a highly conserved Mr 8,000 light chain.
    J Biol Chem. 1996 Aug 9;271(32):19358-66 PMID: 8702622
  16. Tyrosine phosphorylation of I kappa B-alpha activates NF-kappa B without proteolytic degradation of I kappa B-alpha.
    Cell. 1996 Sep 6;86(5):787-98 PMID: 8797825
  17. Role of unphosphorylated, newly synthesized I kappa B beta in persistent activation of NF-kappa B.
    Mol Cell Biol. 1996 Oct;16(10):5444-9 PMID: 8816457
  18. Assembly of focal adhesions: progress, paradigms, and portents.
    Curr Opin Cell Biol. 1996 Feb;8(1):74-85 PMID: 8791409
  19. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.
    Mol Cell Biol. 1985 Dec;5(12):3610-6 PMID: 3915782
  20. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122-6 PMID: 3095828
  21. Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-kappa B transcription factor.
    Cell. 1988 Apr 22;53(2):211-7 PMID: 3129195
  22. Ubiquitin is a component of the microtubule network.
    Proc Natl Acad Sci U S A. 1988 May;85(9):3019-23 PMID: 2834729
  23. I kappa B: a specific inhibitor of the NF-kappa B transcription factor.
    Science. 1988 Oct 28;242(4878):540-6 PMID: 3140380
  24. The lck tyrosine protein kinase interacts with the cytoplasmic tail of the CD4 glycoprotein through its unique amino-terminal domain.
    Cell. 1989 Nov 17;59(4):627-36 PMID: 2582490
  25. NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway.
    Science. 1993 Mar 26;259(5103):1912-5 PMID: 8096091
  26. Mutual regulation of the transcriptional activator NF-kappa B and its inhibitor, I kappa B-alpha.
    Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2532-6 PMID: 8460169
  27. GAL4-I kappa B alpha and GAL4-I kappa B gamma activate transcription by different mechanisms.
    Nucleic Acids Res. 1993 May 11;21(9):2157-63 PMID: 8502557
  28. Disruption of microtubule network in human monocytes induces expression of interleukin-1 but not that of interleukin-6 nor tumor necrosis factor-alpha. Involvement of protein kinase A stimulation.
    J Biol Chem. 1993 Jun 25;268(18):13675-81 PMID: 8099911
  29. Common structural constituents confer I kappa B activity to NF-kappa B p105 and I kappa B/MAD-3.
    EMBO J. 1993 Jul;12(7):2781-8 PMID: 8334994
  30. Cytoskeletal reorganization and TPA differently modify AP-1 to induce the urokinase-type plasminogen activator gene in LLC-PK1 cells.
    Nucleic Acids Res. 1993 Jul 25;21(15):3365-72 PMID: 8346015
  31. Molecular analysis of the microtubule motor dynein.
    Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8769-72 PMID: 8415604
  32. Cytoplasmic dynein plays a role in mammalian mitotic spindle formation.
    J Cell Biol. 1993 Nov;123(4):849-58 PMID: 8227145
  33. High levels of c-rel expression are associated with programmed cell death in the developing avian embryo and in bone marrow cells in vitro.
    Cell. 1993 Dec 3;75(5):899-912 PMID: 8252626
  34. Promoter analysis of the gene encoding the I kappa B-alpha/MAD3 inhibitor of NF-kappa B: positive regulation by members of the rel/NF-kappa B family.
    EMBO J. 1993 Dec 15;12(13):5043-9 PMID: 8262046
  35. The riddle of morphogenesis: a question of solution chemistry or molecular cell engineering?
    Cell. 1993 Dec 31;75(7):1249-52 PMID: 8269508
  36. The I kappa B proteins: members of a multifunctional family.
    Trends Genet. 1993 Dec;9(12):427-33 PMID: 8122310
  37. Disruption of I kappa B alpha regulation by antisense RNA expression leads to malignant transformation.
    Oncogene. 1994 Nov;9(11):3189-97 PMID: 7936641
  38. Subcellular redistribution of HTLV-1 Tax protein by NF-kappa B/Rel transcription factors.
    Virology. 1994 Nov 1;204(2):706-16 PMID: 7941339
  39. A proteasome inhibitor prevents activation of NF-kappa B and stabilizes a newly phosphorylated form of I kappa B-alpha that is still bound to NF-kappa B.
    EMBO J. 1994 Nov 15;13(22):5433-41 PMID: 7957109
  40. Inducible phosphorylation of I kappa B alpha is not sufficient for its dissociation from NF-kappa B and is inhibited by protease inhibitors.
    Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11884-8 PMID: 7991551
  41. Association of p62, a multifunctional SH2- and SH3-domain-binding protein, with src family tyrosine kinases, Grb2, and phospholipase C gamma-1.
    Mol Cell Biol. 1995 Jan;15(1):186-97 PMID: 7799925
  42. Tumor necrosis factor alpha-induced phosphorylation of I kappa B alpha is a signal for its degradation but not dissociation from NF-kappa B.
    Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12740-4 PMID: 7809113
  43. Activation of NF-kappa B requires proteolysis of the inhibitor I kappa B-alpha: signal-induced phosphorylation of I kappa B-alpha alone does not release active NF-kappa B.
    Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):552-6 PMID: 7831327
  44. In vivo stimulation of I kappa B phosphorylation is not sufficient to activate NF-kappa B.
    Mol Cell Biol. 1995 Mar;15(3):1294-301 PMID: 7862123
  45. Phosphorylation of I kappa B alpha precedes but is not sufficient for its dissociation from NF-kappa B.
    Mol Cell Biol. 1995 Mar;15(3):1302-11 PMID: 7862124
  46. Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation.
    Science. 1995 Mar 10;267(5203):1485-8 PMID: 7878466
  47. Domain organization of I kappa B alpha and sites of interaction with NF-kappa B p65.
    Mol Cell Biol. 1995 Apr;15(4):2166-72 PMID: 7891711
  48. Cytoskeletal control of gene expression: depolymerization of microtubules activates NF-kappa B.
    J Cell Biol. 1995 Mar;128(6):1111-9 PMID: 7896875
  49. Activation of the small GTP-binding proteins rho and rac by growth factor receptors.
    J Cell Sci. 1995 Jan;108 ( Pt 1):225-33 PMID: 7738099
  50. Inducible nuclear expression of newly synthesized I kappa B alpha negatively regulates DNA-binding and transcriptional activities of NF-kappa B.
    Mol Cell Biol. 1995 May;15(5):2689-96 PMID: 7739549
  51. Coupling of a signal response domain in I kappa B alpha to multiple pathways for NF-kappa B activation.
    Mol Cell Biol. 1995 May;15(5):2809-18 PMID: 7739562
  52. The M(r) = 8,000 and 11,000 outer arm dynein light chains from Chlamydomonas flagella have cytoplasmic homologues.
    J Biol Chem. 1995 May 12;270(19):11445-52 PMID: 7744782
  53. Identification of a new family of tissue-specific basic helix-loop-helix proteins with a two-hybrid system.
    Mol Cell Biol. 1995 Jul;15(7):3813-22 PMID: 7791788
  54. Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli.
    EMBO J. 1995 Jun 15;14(12):2876-83 PMID: 7796813
  55. Embryonic lethality and liver degeneration in mice lacking the RelA component of NF-kappa B.
    Nature. 1995 Jul 13;376(6536):167-70 PMID: 7603567
  56. HIV Rev uses a conserved cellular protein export pathway for the nucleocytoplasmic transport of viral RNAs.
    Curr Biol. 1996 Jul 1;6(7):848-54 PMID: 8805303
  57. Basal phosphorylation of the PEST domain in the I(kappa)B(beta) regulates its functional interaction with the c-rel proto-oncogene product.
    Mol Cell Biol. 1996 Nov;16(11):5974-84 PMID: 8887627
  58. rac1 regulates a cytokine-stimulated, redox-dependent pathway necessary for NF-kappaB activation.
    Mol Cell Biol. 1996 Dec;16(12):7115-21 PMID: 8943367
  59. Activation of the IkappaB alpha kinase complex by MEKK1, a kinase of the JNK pathway.
    Cell. 1997 Jan 24;88(2):213-22 PMID: 9008162
  60. Nuclear localization of I kappa B alpha promotes active transport of NF-kappa B from the nucleus to the cytoplasm.
    J Cell Sci. 1997 Feb;110 ( Pt 3):369-78 PMID: 9057089
  61. Identification and characterization of an IkappaB kinase.
    Cell. 1997 Jul 25;90(2):373-83 PMID: 9244310
  62. Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway.
    Genes Dev. 1995 Jul 1;9(13):1586-97 PMID: 7628694
  63. Mice lacking the c-rel proto-oncogene exhibit defects in lymphocyte proliferation, humoral immunity, and interleukin-2 expression.
    Genes Dev. 1995 Aug 15;9(16):1965-77 PMID: 7649478
  64. Association of mitogen-activated protein kinase with the microtubule cytoskeleton.
    Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8881-5 PMID: 7568036
  65. Signal-induced degradation of I kappa B alpha requires site-specific ubiquitination.
    Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11259-63 PMID: 7479976
  66. Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation.
    Genes Dev. 1995 Nov 15;9(22):2723-35 PMID: 7590248
  67. Cell polarity. The importance of being polar.
    Curr Biol. 1995 Oct 1;5(10):1102-5 PMID: 8548277
  68. Cytoplasmic dynein is associated with slow axonal transport.
    Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):141-4 PMID: 8552592
  69. Site-specific phosphorylation of IkappaBalpha by a novel ubiquitination-dependent protein kinase activity.
    Cell. 1996 Mar 22;84(6):853-62 PMID: 8601309
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.12.7375
Published
1997-12-00
Pages
7375-85
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232593
Subset
IM
Databases
GENBANK
U00043, U19490, U32855, U32944, U55992
External Links
PubMed source DOI: 10.1128/MCB.17.12.7375 PMC Full Text View journal details
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