CD3 gamma contains a phosphoserine-dependent di-leucine motif involved in down-regulation of the T cell receptor

J Dietrich; X Hou; A M Wegener; C Geisler

CD3 gamma contains a phosphoserine-dependent di-leucine motif involved in down-regulation of the T cell receptor

J Dietrich, X Hou, A M Wegener, C Geisler

Ph.d.-studienævnet for Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

206 Citationer (Scopus)

Abstract

Udgivelsesdato: 1994-May-1

Originalsprog	Engelsk
Tidsskrift	EMBO Journal
Vol/bind	13
Udgave nummer	9
Sider (fra-til)	2156-66
Antal sider	10
ISSN	0261-4189
Status	Udgivet - 1994

Bibliografisk note

Keywords: Amino Acid Sequence; Antigens, CD3; Cell Line; Coated Pits, Cell-Membrane; Down-Regulation; Endocytosis; Enzyme Activation; Humans; Leucine; Molecular Sequence Data; Phosphorylation; Phosphoserine; Point Mutation; Protein Kinase C; Receptors, Antigen, T-Cell; T-Lymphocytes; Transfection; Tyrosine

Citationsformater

@article{da159490b0a611ddb538000ea68e967b,

title = "CD3 gamma contains a phosphoserine-dependent di-leucine motif involved in down-regulation of the T cell receptor",

abstract = "Several cell surface receptors including the T cell receptor (TCR) are phosphorylated and down-regulated following activation of protein kinase C (PKC). Among other substrates the activated PKC in T cells phosphorylates the CD3 gamma subunit of the TCR. To investigate the role of CD3 gamma phosphorylation in PKC-mediated TCR down-regulation, point mutated CD3 gamma cDNA was transfected into the CD3 gamma-negative T cell line JGN and CD3 gamma transfectants were analysed. Phosphorylation at S126 but not S123 in the cytoplasmic tail of CD3 gamma was required for PKC-mediated down-regulation of the TCR. Furthermore, analysis of a series of CD3 gamma truncation mutants indicated that in addition to S126 phosphorylation a motif C-terminal of S126 was required for TCR down-regulation. Point mutation analyses confirmed this observation and demonstrated that a membrane-proximal di-leucine motif (L131 and L132) in the cytoplasmic tail of CD3 gamma was required for PKC-mediated TCR down-regulation in addition to phosphorylation at S126. Incubation of T cells in hypertonic medium known to disrupt normal clathrin lattices severely inhibited PKC-mediated TCR down-regulation in non-mutated T cells, indicating that the TCR was down-regulated by endocytosis via clathrin coated pits. Based on the present results and previously published observations on intracellular receptor sorting, a general model for intracellular sorting of receptors containing di-leucine- or tyrosine-based motifs is proposed.",

author = "J Dietrich and X Hou and Wegener, {A M} and C Geisler",

note = "Keywords: Amino Acid Sequence; Antigens, CD3; Cell Line; Coated Pits, Cell-Membrane; Down-Regulation; Endocytosis; Enzyme Activation; Humans; Leucine; Molecular Sequence Data; Phosphorylation; Phosphoserine; Point Mutation; Protein Kinase C; Receptors, Antigen, T-Cell; T-Lymphocytes; Transfection; Tyrosine",

year = "1994",

language = "English",

volume = "13",

pages = "2156--66",

journal = "EMBO Journal",

issn = "0261-4189",

publisher = "Wiley-Blackwell",

number = "9",

}

TY - JOUR

T1 - CD3 gamma contains a phosphoserine-dependent di-leucine motif involved in down-regulation of the T cell receptor

AU - Dietrich, J

AU - Hou, X

AU - Wegener, A M

AU - Geisler, C

N1 - Keywords: Amino Acid Sequence; Antigens, CD3; Cell Line; Coated Pits, Cell-Membrane; Down-Regulation; Endocytosis; Enzyme Activation; Humans; Leucine; Molecular Sequence Data; Phosphorylation; Phosphoserine; Point Mutation; Protein Kinase C; Receptors, Antigen, T-Cell; T-Lymphocytes; Transfection; Tyrosine

PY - 1994

Y1 - 1994

N2 - Several cell surface receptors including the T cell receptor (TCR) are phosphorylated and down-regulated following activation of protein kinase C (PKC). Among other substrates the activated PKC in T cells phosphorylates the CD3 gamma subunit of the TCR. To investigate the role of CD3 gamma phosphorylation in PKC-mediated TCR down-regulation, point mutated CD3 gamma cDNA was transfected into the CD3 gamma-negative T cell line JGN and CD3 gamma transfectants were analysed. Phosphorylation at S126 but not S123 in the cytoplasmic tail of CD3 gamma was required for PKC-mediated down-regulation of the TCR. Furthermore, analysis of a series of CD3 gamma truncation mutants indicated that in addition to S126 phosphorylation a motif C-terminal of S126 was required for TCR down-regulation. Point mutation analyses confirmed this observation and demonstrated that a membrane-proximal di-leucine motif (L131 and L132) in the cytoplasmic tail of CD3 gamma was required for PKC-mediated TCR down-regulation in addition to phosphorylation at S126. Incubation of T cells in hypertonic medium known to disrupt normal clathrin lattices severely inhibited PKC-mediated TCR down-regulation in non-mutated T cells, indicating that the TCR was down-regulated by endocytosis via clathrin coated pits. Based on the present results and previously published observations on intracellular receptor sorting, a general model for intracellular sorting of receptors containing di-leucine- or tyrosine-based motifs is proposed.

AB - Several cell surface receptors including the T cell receptor (TCR) are phosphorylated and down-regulated following activation of protein kinase C (PKC). Among other substrates the activated PKC in T cells phosphorylates the CD3 gamma subunit of the TCR. To investigate the role of CD3 gamma phosphorylation in PKC-mediated TCR down-regulation, point mutated CD3 gamma cDNA was transfected into the CD3 gamma-negative T cell line JGN and CD3 gamma transfectants were analysed. Phosphorylation at S126 but not S123 in the cytoplasmic tail of CD3 gamma was required for PKC-mediated down-regulation of the TCR. Furthermore, analysis of a series of CD3 gamma truncation mutants indicated that in addition to S126 phosphorylation a motif C-terminal of S126 was required for TCR down-regulation. Point mutation analyses confirmed this observation and demonstrated that a membrane-proximal di-leucine motif (L131 and L132) in the cytoplasmic tail of CD3 gamma was required for PKC-mediated TCR down-regulation in addition to phosphorylation at S126. Incubation of T cells in hypertonic medium known to disrupt normal clathrin lattices severely inhibited PKC-mediated TCR down-regulation in non-mutated T cells, indicating that the TCR was down-regulated by endocytosis via clathrin coated pits. Based on the present results and previously published observations on intracellular receptor sorting, a general model for intracellular sorting of receptors containing di-leucine- or tyrosine-based motifs is proposed.

M3 - Journal article

C2 - 8187769

SN - 0261-4189

VL - 13

SP - 2156

EP - 2166

JO - EMBO Journal

JF - EMBO Journal

IS - 9

ER -