Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells

M Lub, S J van Vliet, S P Oomen, R A Pieters, M Robinson, C G Figdor, Y van Kooyk

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The beta 2 integrin lymphocyte function-associated antigen 1 (LFA-1) mediates activation-dependent adhesion of lymphocytes. To investigate whether lymphocyte-specific elements are essential for LFA-1 function, we expressed LFA-1 in the erythroleukemic cell line K562, which expresses only the integrin very late antigen 5. We observed that LFA-1-expressing K562 cannot bind to intercellular adhesion molecule 1-coated surfaces when stimulated by phorbol 12-myristate 13-acetate (PMA), whereas the LFA-1-activating antibody KIM185 markedly enhanced adhesion. Because the endogenously expressed beta 1 integrin very late antigen 5 is readily activated by PMA, we investigated the role of the cytoplasmic domain of distinct beta subunits in regulating LFA-1 function. Transfection of chimeric LFA-1 receptors in K562 cells reveals that replacement of the beta 2 cytoplasmic tail with the beta 1 but not the beta 7 cytoplasmic tail completely restores PMA responsiveness of LFA-1, whereas a beta 2 cytoplasmic deletion mutant of LFA-1 is constitutively active. Both deletion of the beta 2 cytoplasmic tail or replacement by the beta 1 cytoplasmic tail alters the localization of LFA-1 into clusters, thereby regulating LFA-1 activation and LFA-1-mediated adhesion to intercellular adhesion molecule 1. These data demonstrate that distinct signaling routes activate beta 1 and beta 2 integrins through the beta-chain and hint at the involvement of lymphocyte-specific signal transduction elements in beta 2 and beta 7 integrin activation that are absent in the nonlymphocytic cell line K562.

Original languageEnglish
Pages (from-to)719-28
Number of pages10
JournalMolecular Biology of the Cell
Volume8
Issue number4
Publication statusPublished - Apr 1997

Cite this

Lub, M., van Vliet, S. J., Oomen, S. P., Pieters, R. A., Robinson, M., Figdor, C. G., & van Kooyk, Y. (1997). Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells. Molecular Biology of the Cell, 8(4), 719-28.
Lub, M ; van Vliet, S J ; Oomen, S P ; Pieters, R A ; Robinson, M ; Figdor, C G ; van Kooyk, Y. / Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells. In: Molecular Biology of the Cell. 1997 ; Vol. 8, No. 4. pp. 719-28.
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abstract = "The beta 2 integrin lymphocyte function-associated antigen 1 (LFA-1) mediates activation-dependent adhesion of lymphocytes. To investigate whether lymphocyte-specific elements are essential for LFA-1 function, we expressed LFA-1 in the erythroleukemic cell line K562, which expresses only the integrin very late antigen 5. We observed that LFA-1-expressing K562 cannot bind to intercellular adhesion molecule 1-coated surfaces when stimulated by phorbol 12-myristate 13-acetate (PMA), whereas the LFA-1-activating antibody KIM185 markedly enhanced adhesion. Because the endogenously expressed beta 1 integrin very late antigen 5 is readily activated by PMA, we investigated the role of the cytoplasmic domain of distinct beta subunits in regulating LFA-1 function. Transfection of chimeric LFA-1 receptors in K562 cells reveals that replacement of the beta 2 cytoplasmic tail with the beta 1 but not the beta 7 cytoplasmic tail completely restores PMA responsiveness of LFA-1, whereas a beta 2 cytoplasmic deletion mutant of LFA-1 is constitutively active. Both deletion of the beta 2 cytoplasmic tail or replacement by the beta 1 cytoplasmic tail alters the localization of LFA-1 into clusters, thereby regulating LFA-1 activation and LFA-1-mediated adhesion to intercellular adhesion molecule 1. These data demonstrate that distinct signaling routes activate beta 1 and beta 2 integrins through the beta-chain and hint at the involvement of lymphocyte-specific signal transduction elements in beta 2 and beta 7 integrin activation that are absent in the nonlymphocytic cell line K562.",
keywords = "Amino Acid Sequence, Binding Sites, CD18 Antigens/drug effects, Cell Adhesion/physiology, Cell Membrane/metabolism, Cytoplasm/metabolism, Fibronectins/metabolism, Humans, Integrin beta Chains, Integrin beta1/drug effects, Integrins/drug effects, Intercellular Adhesion Molecule-1/metabolism, Leukemia, Erythroblastic, Acute/drug therapy, Leukemia, Experimental, Lymphocyte Function-Associated Antigen-1/drug effects, Lymphocytes/physiology, Molecular Sequence Data, Recombinant Proteins/drug effects, Signal Transduction, Tetradecanoylphorbol Acetate/pharmacology, Tumor Cells, Cultured",
author = "M Lub and {van Vliet}, {S J} and Oomen, {S P} and Pieters, {R A} and M Robinson and Figdor, {C G} and {van Kooyk}, Y",
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Lub, M, van Vliet, SJ, Oomen, SP, Pieters, RA, Robinson, M, Figdor, CG & van Kooyk, Y 1997, 'Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells' Molecular Biology of the Cell, vol. 8, no. 4, pp. 719-28.

Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells. / Lub, M; van Vliet, S J; Oomen, S P; Pieters, R A; Robinson, M; Figdor, C G; van Kooyk, Y.

In: Molecular Biology of the Cell, Vol. 8, No. 4, 04.1997, p. 719-28.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells

AU - Lub, M

AU - van Vliet, S J

AU - Oomen, S P

AU - Pieters, R A

AU - Robinson, M

AU - Figdor, C G

AU - van Kooyk, Y

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N2 - The beta 2 integrin lymphocyte function-associated antigen 1 (LFA-1) mediates activation-dependent adhesion of lymphocytes. To investigate whether lymphocyte-specific elements are essential for LFA-1 function, we expressed LFA-1 in the erythroleukemic cell line K562, which expresses only the integrin very late antigen 5. We observed that LFA-1-expressing K562 cannot bind to intercellular adhesion molecule 1-coated surfaces when stimulated by phorbol 12-myristate 13-acetate (PMA), whereas the LFA-1-activating antibody KIM185 markedly enhanced adhesion. Because the endogenously expressed beta 1 integrin very late antigen 5 is readily activated by PMA, we investigated the role of the cytoplasmic domain of distinct beta subunits in regulating LFA-1 function. Transfection of chimeric LFA-1 receptors in K562 cells reveals that replacement of the beta 2 cytoplasmic tail with the beta 1 but not the beta 7 cytoplasmic tail completely restores PMA responsiveness of LFA-1, whereas a beta 2 cytoplasmic deletion mutant of LFA-1 is constitutively active. Both deletion of the beta 2 cytoplasmic tail or replacement by the beta 1 cytoplasmic tail alters the localization of LFA-1 into clusters, thereby regulating LFA-1 activation and LFA-1-mediated adhesion to intercellular adhesion molecule 1. These data demonstrate that distinct signaling routes activate beta 1 and beta 2 integrins through the beta-chain and hint at the involvement of lymphocyte-specific signal transduction elements in beta 2 and beta 7 integrin activation that are absent in the nonlymphocytic cell line K562.

AB - The beta 2 integrin lymphocyte function-associated antigen 1 (LFA-1) mediates activation-dependent adhesion of lymphocytes. To investigate whether lymphocyte-specific elements are essential for LFA-1 function, we expressed LFA-1 in the erythroleukemic cell line K562, which expresses only the integrin very late antigen 5. We observed that LFA-1-expressing K562 cannot bind to intercellular adhesion molecule 1-coated surfaces when stimulated by phorbol 12-myristate 13-acetate (PMA), whereas the LFA-1-activating antibody KIM185 markedly enhanced adhesion. Because the endogenously expressed beta 1 integrin very late antigen 5 is readily activated by PMA, we investigated the role of the cytoplasmic domain of distinct beta subunits in regulating LFA-1 function. Transfection of chimeric LFA-1 receptors in K562 cells reveals that replacement of the beta 2 cytoplasmic tail with the beta 1 but not the beta 7 cytoplasmic tail completely restores PMA responsiveness of LFA-1, whereas a beta 2 cytoplasmic deletion mutant of LFA-1 is constitutively active. Both deletion of the beta 2 cytoplasmic tail or replacement by the beta 1 cytoplasmic tail alters the localization of LFA-1 into clusters, thereby regulating LFA-1 activation and LFA-1-mediated adhesion to intercellular adhesion molecule 1. These data demonstrate that distinct signaling routes activate beta 1 and beta 2 integrins through the beta-chain and hint at the involvement of lymphocyte-specific signal transduction elements in beta 2 and beta 7 integrin activation that are absent in the nonlymphocytic cell line K562.

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KW - Binding Sites

KW - CD18 Antigens/drug effects

KW - Cell Adhesion/physiology

KW - Cell Membrane/metabolism

KW - Cytoplasm/metabolism

KW - Fibronectins/metabolism

KW - Humans

KW - Integrin beta Chains

KW - Integrin beta1/drug effects

KW - Integrins/drug effects

KW - Intercellular Adhesion Molecule-1/metabolism

KW - Leukemia, Erythroblastic, Acute/drug therapy

KW - Leukemia, Experimental

KW - Lymphocyte Function-Associated Antigen-1/drug effects

KW - Lymphocytes/physiology

KW - Molecular Sequence Data

KW - Recombinant Proteins/drug effects

KW - Signal Transduction

KW - Tetradecanoylphorbol Acetate/pharmacology

KW - Tumor Cells, Cultured

M3 - Article

VL - 8

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EP - 728

JO - Molecular Biology of the Cell

JF - Molecular Biology of the Cell

SN - 1059-1524

IS - 4

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Lub M, van Vliet SJ, Oomen SP, Pieters RA, Robinson M, Figdor CG et al. Cytoplasmic tails of beta 1, beta 2, and beta 7 integrins differentially regulate LFA-1 function in K562 cells. Molecular Biology of the Cell. 1997 Apr;8(4):719-28.