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PMID: 11904433 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Negative regulation of Lck by Cbl ubiquitin ligase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 99 ·No. 6 ·2002-03-19 ·Pages 3794-9

Rao N, Miyake S, Reddi AL, Douillard P, Ghosh AK, Dodge IL, Zhou P, Fernandes ND, Band H

Abstract

The Cbl-family ubiquitin ligases function as negative regulators of activated receptor tyrosine kinases by facilitating their ubiquitination and subsequent targeting to lysosomes. Cbl associates with the lymphoid-restricted nonreceptor tyrosine kinase Lck, but the functional relevance of this interaction remains unknown. Here, we demonstrate that T cell receptor and CD4 coligation on human T cells results in enhanced association between Cbl and Lck, together with Lck ubiquitination and degradation. A Cbl(-/-) T cell line showed a marked deficiency in Lck ubiquitination and increased levels of kinase-active Lck. Coexpression in 293T cells demonstrated that Lck kinase activity and Cbl ubiquitin ligase activity were essential for Lck ubiquitination and negative regulation of Lck-dependent serum response element-luciferase reporter activity. The Lck SH3 domain was pivotal for Cbl-Lck association and Cbl-mediated Lck degradation, with a smaller role for interactions mediated by the Cbl tyrosine kinase-binding domain. Finally, analysis of a ZAP-70-deficient T cell line revealed that Cbl inhibited Lck-dependent mitogen-activated protein kinase activation, and an intact Cbl RING finger domain was required for this functional effect. Our results demonstrate a direct, ubiquitination-dependent, negative regulatory role of Cbl for Lck in T cells, independent of Cbl-mediated regulation of ZAP-70.

MeSH Terms
CD4 Antigens/metabolism Cell Line Enzyme Activation Gene Deletion Genes, Reporter/genetics Humans Jurkat Cells Ligases/chemistry,deficiency,genetics,metabolism Lymphocyte Activation Lymphocyte Specific Protein Tyrosine Kinase p56(lck)/antagonists & inhibitors,chemistry,genetics,metabolism Protein Binding Protein Processing, Post-Translational Protein Structure, Tertiary Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/chemistry,deficiency,genetics,metabolism Proto-Oncogene Proteins c-cbl Receptors, Antigen, T-Cell/metabolism T-Lymphocytes/immunology,metabolism Ubiquitin/metabolism Ubiquitin-Protein Ligases ZAP-70 Protein-Tyrosine Kinase src Homology Domains
Chemicals
CD4 Antigens Proto-Oncogene Proteins Receptors, Antigen, T-Cell Ubiquitin Proto-Oncogene Proteins c-cbl Ubiquitin-Protein Ligases Protein-Tyrosine Kinases Lymphocyte Specific Protein Tyrosine Kinase p56(lck) ZAP-70 Protein-Tyrosine Kinase ZAP70 protein, human Ligases CBL protein, human
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Rao Navin
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Miyake Sachiko
Reddi Alagarsamy Lakku
Douillard Patrice
Ghosh Amiya K
Dodge Ingrid L
Zhou Pengcheng
Fernandes Norvin D
Band Hamid
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39 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.062055999
Published
2002-03-19
Pages
3794-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC122603
Subset
IM
Grants
NCI NIH HHS · R01 CA087986 · United States
NIAMS NIH HHS · T32 AR007530 · United States
NCI NIH HHS · CA75075 · United States
NCI NIH HHS · CA76118 · United States
NIAMS NIH HHS · 5T32AR07530 · United States
NCI NIH HHS · CA87986 · United States
External Links
PubMed source DOI: 10.1073/pnas.062055999 PMC Full Text View journal details
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