The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation

Navin Rao, Amiya K. Ghosh, Satoshi Ota, Pengcheng Zhou, Alagarsamy Lakku Reddi, Kaoru Hakezi, Brian Druker, Jiong Wu, Hamid Band

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

The negative regulator Cbl functions as a ubiquitin ligase towards activated receptor tyrosine kinases and facilitates their transport to lysosomes. Whether Cbl ubiquitin ligase activity mediates its negative regulatory effects on cytoplasmic tyrosine kinases of the Syk/ZAP-70 family has not been addressed, nor is it known whether these kinases are regulated via ubiquitylation during lymphocyte B-cell receptor engagement. Here we show that B-cell receptor stimulation in Ramos cells induces the ubiquitylation of Syk tyrosine kinase which is inhibited by a dominant-negative mutant of Cbl. Intact tyrosine kinase-binding and RING finger domains of Cbl were found to be essential for Syk ubiquitylation in 293T cells and for in vitro Syk ubiquitylation. These same domains were also essential for Cbl-mediated negative regulation of Syk as measured using an NFAT-luciferase reporter in a lymphoid cell. Association with Cbl did not alter the kinase activity of Syk. Altogether, our results support an essential role for Cbl ubiquitin ligase activity in the negative regulation of Syk, and establish that ubiquitylation provides a mechanism of Cbl-mediated negative regulation of cytoplasmic targets.

Original languageEnglish (US)
Pages (from-to)7085-7095
Number of pages11
JournalEMBO Journal
Volume20
Issue number24
DOIs
StatePublished - Dec 17 2002

Fingerprint

Ubiquitination
Ligases
Ubiquitin
Protein-Tyrosine Kinases
B-Lymphocytes
Cells
Phosphotransferases
Lymphocytes
Receptor Protein-Tyrosine Kinases
Luciferases
RING Finger Domains
HEK293 Cells
Lysosomes
Syk Kinase

Keywords

  • Activation
  • Lymphocyte
  • Regulation
  • Tyrosine kinase
  • Ubiquitylation

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Rao, N., Ghosh, A. K., Ota, S., Zhou, P., Reddi, A. L., Hakezi, K., ... Band, H. (2002). The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation. EMBO Journal, 20(24), 7085-7095. https://doi.org/10.1093/emboj/20.24.7085

The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation. / Rao, Navin; Ghosh, Amiya K.; Ota, Satoshi; Zhou, Pengcheng; Reddi, Alagarsamy Lakku; Hakezi, Kaoru; Druker, Brian; Wu, Jiong; Band, Hamid.

In: EMBO Journal, Vol. 20, No. 24, 17.12.2002, p. 7085-7095.

Research output: Contribution to journalArticle

Rao, N, Ghosh, AK, Ota, S, Zhou, P, Reddi, AL, Hakezi, K, Druker, B, Wu, J & Band, H 2002, 'The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation', EMBO Journal, vol. 20, no. 24, pp. 7085-7095. https://doi.org/10.1093/emboj/20.24.7085
Rao, Navin ; Ghosh, Amiya K. ; Ota, Satoshi ; Zhou, Pengcheng ; Reddi, Alagarsamy Lakku ; Hakezi, Kaoru ; Druker, Brian ; Wu, Jiong ; Band, Hamid. / The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation. In: EMBO Journal. 2002 ; Vol. 20, No. 24. pp. 7085-7095.
@article{cefae0ade58d45e09872a9578006bba6,
title = "The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation",
abstract = "The negative regulator Cbl functions as a ubiquitin ligase towards activated receptor tyrosine kinases and facilitates their transport to lysosomes. Whether Cbl ubiquitin ligase activity mediates its negative regulatory effects on cytoplasmic tyrosine kinases of the Syk/ZAP-70 family has not been addressed, nor is it known whether these kinases are regulated via ubiquitylation during lymphocyte B-cell receptor engagement. Here we show that B-cell receptor stimulation in Ramos cells induces the ubiquitylation of Syk tyrosine kinase which is inhibited by a dominant-negative mutant of Cbl. Intact tyrosine kinase-binding and RING finger domains of Cbl were found to be essential for Syk ubiquitylation in 293T cells and for in vitro Syk ubiquitylation. These same domains were also essential for Cbl-mediated negative regulation of Syk as measured using an NFAT-luciferase reporter in a lymphoid cell. Association with Cbl did not alter the kinase activity of Syk. Altogether, our results support an essential role for Cbl ubiquitin ligase activity in the negative regulation of Syk, and establish that ubiquitylation provides a mechanism of Cbl-mediated negative regulation of cytoplasmic targets.",
keywords = "Activation, Lymphocyte, Regulation, Tyrosine kinase, Ubiquitylation",
author = "Navin Rao and Ghosh, {Amiya K.} and Satoshi Ota and Pengcheng Zhou and Reddi, {Alagarsamy Lakku} and Kaoru Hakezi and Brian Druker and Jiong Wu and Hamid Band",
year = "2002",
month = "12",
day = "17",
doi = "10.1093/emboj/20.24.7085",
language = "English (US)",
volume = "20",
pages = "7085--7095",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "24",

}

TY - JOUR

T1 - The non-receptor tyrosine kinase Syk is a target of Cbl-mediated ubiquitylation upon B-cell receptor stimulation

AU - Rao, Navin

AU - Ghosh, Amiya K.

AU - Ota, Satoshi

AU - Zhou, Pengcheng

AU - Reddi, Alagarsamy Lakku

AU - Hakezi, Kaoru

AU - Druker, Brian

AU - Wu, Jiong

AU - Band, Hamid

PY - 2002/12/17

Y1 - 2002/12/17

N2 - The negative regulator Cbl functions as a ubiquitin ligase towards activated receptor tyrosine kinases and facilitates their transport to lysosomes. Whether Cbl ubiquitin ligase activity mediates its negative regulatory effects on cytoplasmic tyrosine kinases of the Syk/ZAP-70 family has not been addressed, nor is it known whether these kinases are regulated via ubiquitylation during lymphocyte B-cell receptor engagement. Here we show that B-cell receptor stimulation in Ramos cells induces the ubiquitylation of Syk tyrosine kinase which is inhibited by a dominant-negative mutant of Cbl. Intact tyrosine kinase-binding and RING finger domains of Cbl were found to be essential for Syk ubiquitylation in 293T cells and for in vitro Syk ubiquitylation. These same domains were also essential for Cbl-mediated negative regulation of Syk as measured using an NFAT-luciferase reporter in a lymphoid cell. Association with Cbl did not alter the kinase activity of Syk. Altogether, our results support an essential role for Cbl ubiquitin ligase activity in the negative regulation of Syk, and establish that ubiquitylation provides a mechanism of Cbl-mediated negative regulation of cytoplasmic targets.

AB - The negative regulator Cbl functions as a ubiquitin ligase towards activated receptor tyrosine kinases and facilitates their transport to lysosomes. Whether Cbl ubiquitin ligase activity mediates its negative regulatory effects on cytoplasmic tyrosine kinases of the Syk/ZAP-70 family has not been addressed, nor is it known whether these kinases are regulated via ubiquitylation during lymphocyte B-cell receptor engagement. Here we show that B-cell receptor stimulation in Ramos cells induces the ubiquitylation of Syk tyrosine kinase which is inhibited by a dominant-negative mutant of Cbl. Intact tyrosine kinase-binding and RING finger domains of Cbl were found to be essential for Syk ubiquitylation in 293T cells and for in vitro Syk ubiquitylation. These same domains were also essential for Cbl-mediated negative regulation of Syk as measured using an NFAT-luciferase reporter in a lymphoid cell. Association with Cbl did not alter the kinase activity of Syk. Altogether, our results support an essential role for Cbl ubiquitin ligase activity in the negative regulation of Syk, and establish that ubiquitylation provides a mechanism of Cbl-mediated negative regulation of cytoplasmic targets.

KW - Activation

KW - Lymphocyte

KW - Regulation

KW - Tyrosine kinase

KW - Ubiquitylation

UR - http://www.scopus.com/inward/record.url?scp=0037126634&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037126634&partnerID=8YFLogxK

U2 - 10.1093/emboj/20.24.7085

DO - 10.1093/emboj/20.24.7085

M3 - Article

VL - 20

SP - 7085

EP - 7095

JO - EMBO Journal

JF - EMBO Journal

SN - 0261-4189

IS - 24

ER -