Expression cloning of a lymphocyte homing receptor cDNA: Ubiquitin is the reactive species

Tom St John, W. Michael Gallatin, Mark Siegelman, Harry T. Smith, Victor A. Fried, Irving L. Weissman

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

The lymphocyte cell surface receptor for the high endothelial venules (HEV's) of peripheral lymph nodes is specifically recognized by the monoclonal antibody MEL14. Three independent complementary DNA (cDNA) clones, each of which encodes the protein ubiquitin, were detected by virtue of the expression of the MEL-14 antigenic determinant on cDNA-β-galactosidase bacterial fusion proteins. The antigenic determinant defined by MEL-14 resides in the carboxyl terminal 13-amino-acid proteotytic peptide of ubiquitin, but is undetected in intact undenatured ubiquitin and other cellular ubiquitinated proteins. Antisera and monoclonal antibodies to ubiquitin determinants bind to the surface of both HEV-receptor positive and negative cell lines. The MEL-14-identified cDNA clones hydridize to RNA transcripts that encode tandemly repeated ubiquitins. Sequence analysis of these polyubiquitin cDNA's does not identify a leader sequence for export to the cell surface. The expression of the MEL-14 epitope of ubiquitin depends upon its local environment. The steady-state levels of expression of the ubiquitin messenger RNA's do not correlate with either the tissue derivation of the RNA or the expression of the lymphocyte HEV receptor. Regulation of the expression of the HEV receptor is not likely to reflect the transcriptional control of ubiquitin genes, but rather to reflect control of the expression of the HEV core polypcptide or its level or form of ubiquitination.

Original languageEnglish (US)
Pages (from-to)845-850
Number of pages6
JournalScience
Volume231
Issue number4740
StatePublished - 1986

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Lymphocyte Homing Receptors
Ubiquitin
Organism Cloning
Venules
Complementary DNA
Epitopes
Clone Cells
Monoclonal Antibodies
Ubiquitins
Galactosidases
Ubiquitinated Proteins
Polyubiquitin
Lymphocytes
RNA
Bacterial Proteins
Ubiquitination
Cell Surface Receptors
Sequence Analysis
Immune Sera
Lymph Nodes

ASJC Scopus subject areas

  • General

Cite this

John, T. S., Gallatin, W. M., Siegelman, M., Smith, H. T., Fried, V. A., & Weissman, I. L. (1986). Expression cloning of a lymphocyte homing receptor cDNA: Ubiquitin is the reactive species. Science, 231(4740), 845-850.

Expression cloning of a lymphocyte homing receptor cDNA : Ubiquitin is the reactive species. / John, Tom St; Gallatin, W. Michael; Siegelman, Mark; Smith, Harry T.; Fried, Victor A.; Weissman, Irving L.

In: Science, Vol. 231, No. 4740, 1986, p. 845-850.

Research output: Contribution to journalArticle

John, TS, Gallatin, WM, Siegelman, M, Smith, HT, Fried, VA & Weissman, IL 1986, 'Expression cloning of a lymphocyte homing receptor cDNA: Ubiquitin is the reactive species', Science, vol. 231, no. 4740, pp. 845-850.
John TS, Gallatin WM, Siegelman M, Smith HT, Fried VA, Weissman IL. Expression cloning of a lymphocyte homing receptor cDNA: Ubiquitin is the reactive species. Science. 1986;231(4740):845-850.
John, Tom St ; Gallatin, W. Michael ; Siegelman, Mark ; Smith, Harry T. ; Fried, Victor A. ; Weissman, Irving L. / Expression cloning of a lymphocyte homing receptor cDNA : Ubiquitin is the reactive species. In: Science. 1986 ; Vol. 231, No. 4740. pp. 845-850.
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