Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells

David L. Simmons, Anne B. Satterthwaite, Daniel G. Tenen, Brian Seed

Research output: Contribution to journalArticle

194 Citations (Scopus)

Abstract

CD34 is a 115-kDa transmembrane glycoprotein of unknown function that is expressed on human hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues. We have isolated a CD34 cDNA clone from a KG-1 cell library following three rounds of transient expression in COS cells and enrichment by panning with the anti-CD34 mAb MY10 and BI-3C5. The 5′ and 3′ ends of the full-length cDNA were subsequently amplified by polymerase chain reaction from KG-1 RNA; the final cDNA clone contained 2615 bp and ended in a poly(A) tail. COS cells transfected with the cDNA clone expressed a surface protein of approximately 110 kDa that was immunoprecipitated by MY10. Southern blot analysis suggested that CD34 is a single copy gene. A 2.7-kb CD34 transcript was observed in the hematopoietic cell lines KG-1, KMT-2, AML-1, RPMI 8402, and MOLT 13 and the endothelial cells BAE and EAhy926, but not in monocytes, resting T cells, or the cell lines Laz 509, HL-60, U937, K562, and HeLa. The cDNA sequence predicts a 40-kDa type I integral membrane protein with nine potential N-linked and numerous potential O-linked glycosylation sites in its extracellular domain. There are two consensus protein kinase C phosphorylation sites and one potential tyrosine kinase phosphorylation site in the cytoplasmic portion of CD34. CD34 has no significant sequence homology to any known protein but has some structural similarities to the heavily glycosylated leukocyte surface molecule CD43.

Original languageEnglish (US)
Pages (from-to)267-271
Number of pages5
JournalJournal of Immunology
Volume148
Issue number1
StatePublished - Jan 1 1992

Fingerprint

Sialomucins
Molecular Cloning
Hematopoietic Stem Cells
Complementary DNA
Clone Cells
COS Cells
Membrane Proteins
Phosphorylation
Cell Line
Sequence Homology
Southern Blotting
Glycosylation
Protein-Tyrosine Kinases
Protein Kinase C
Endothelium
Monocytes
Glycoproteins
Leukocytes
Endothelial Cells
RNA

ASJC Scopus subject areas

  • Immunology

Cite this

Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells. / Simmons, David L.; Satterthwaite, Anne B.; Tenen, Daniel G.; Seed, Brian.

In: Journal of Immunology, Vol. 148, No. 1, 01.01.1992, p. 267-271.

Research output: Contribution to journalArticle

Simmons, David L. ; Satterthwaite, Anne B. ; Tenen, Daniel G. ; Seed, Brian. / Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells. In: Journal of Immunology. 1992 ; Vol. 148, No. 1. pp. 267-271.
@article{29ec7122b79743c390e41c0840a129ae,
title = "Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells",
abstract = "CD34 is a 115-kDa transmembrane glycoprotein of unknown function that is expressed on human hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues. We have isolated a CD34 cDNA clone from a KG-1 cell library following three rounds of transient expression in COS cells and enrichment by panning with the anti-CD34 mAb MY10 and BI-3C5. The 5′ and 3′ ends of the full-length cDNA were subsequently amplified by polymerase chain reaction from KG-1 RNA; the final cDNA clone contained 2615 bp and ended in a poly(A) tail. COS cells transfected with the cDNA clone expressed a surface protein of approximately 110 kDa that was immunoprecipitated by MY10. Southern blot analysis suggested that CD34 is a single copy gene. A 2.7-kb CD34 transcript was observed in the hematopoietic cell lines KG-1, KMT-2, AML-1, RPMI 8402, and MOLT 13 and the endothelial cells BAE and EAhy926, but not in monocytes, resting T cells, or the cell lines Laz 509, HL-60, U937, K562, and HeLa. The cDNA sequence predicts a 40-kDa type I integral membrane protein with nine potential N-linked and numerous potential O-linked glycosylation sites in its extracellular domain. There are two consensus protein kinase C phosphorylation sites and one potential tyrosine kinase phosphorylation site in the cytoplasmic portion of CD34. CD34 has no significant sequence homology to any known protein but has some structural similarities to the heavily glycosylated leukocyte surface molecule CD43.",
author = "Simmons, {David L.} and Satterthwaite, {Anne B.} and Tenen, {Daniel G.} and Brian Seed",
year = "1992",
month = "1",
day = "1",
language = "English (US)",
volume = "148",
pages = "267--271",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "1",

}

TY - JOUR

T1 - Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells

AU - Simmons, David L.

AU - Satterthwaite, Anne B.

AU - Tenen, Daniel G.

AU - Seed, Brian

PY - 1992/1/1

Y1 - 1992/1/1

N2 - CD34 is a 115-kDa transmembrane glycoprotein of unknown function that is expressed on human hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues. We have isolated a CD34 cDNA clone from a KG-1 cell library following three rounds of transient expression in COS cells and enrichment by panning with the anti-CD34 mAb MY10 and BI-3C5. The 5′ and 3′ ends of the full-length cDNA were subsequently amplified by polymerase chain reaction from KG-1 RNA; the final cDNA clone contained 2615 bp and ended in a poly(A) tail. COS cells transfected with the cDNA clone expressed a surface protein of approximately 110 kDa that was immunoprecipitated by MY10. Southern blot analysis suggested that CD34 is a single copy gene. A 2.7-kb CD34 transcript was observed in the hematopoietic cell lines KG-1, KMT-2, AML-1, RPMI 8402, and MOLT 13 and the endothelial cells BAE and EAhy926, but not in monocytes, resting T cells, or the cell lines Laz 509, HL-60, U937, K562, and HeLa. The cDNA sequence predicts a 40-kDa type I integral membrane protein with nine potential N-linked and numerous potential O-linked glycosylation sites in its extracellular domain. There are two consensus protein kinase C phosphorylation sites and one potential tyrosine kinase phosphorylation site in the cytoplasmic portion of CD34. CD34 has no significant sequence homology to any known protein but has some structural similarities to the heavily glycosylated leukocyte surface molecule CD43.

AB - CD34 is a 115-kDa transmembrane glycoprotein of unknown function that is expressed on human hematopoietic progenitor cells and the small vessel endothelium of a variety of tissues. We have isolated a CD34 cDNA clone from a KG-1 cell library following three rounds of transient expression in COS cells and enrichment by panning with the anti-CD34 mAb MY10 and BI-3C5. The 5′ and 3′ ends of the full-length cDNA were subsequently amplified by polymerase chain reaction from KG-1 RNA; the final cDNA clone contained 2615 bp and ended in a poly(A) tail. COS cells transfected with the cDNA clone expressed a surface protein of approximately 110 kDa that was immunoprecipitated by MY10. Southern blot analysis suggested that CD34 is a single copy gene. A 2.7-kb CD34 transcript was observed in the hematopoietic cell lines KG-1, KMT-2, AML-1, RPMI 8402, and MOLT 13 and the endothelial cells BAE and EAhy926, but not in monocytes, resting T cells, or the cell lines Laz 509, HL-60, U937, K562, and HeLa. The cDNA sequence predicts a 40-kDa type I integral membrane protein with nine potential N-linked and numerous potential O-linked glycosylation sites in its extracellular domain. There are two consensus protein kinase C phosphorylation sites and one potential tyrosine kinase phosphorylation site in the cytoplasmic portion of CD34. CD34 has no significant sequence homology to any known protein but has some structural similarities to the heavily glycosylated leukocyte surface molecule CD43.

UR - http://www.scopus.com/inward/record.url?scp=0026515407&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026515407&partnerID=8YFLogxK

M3 - Article

C2 - 1370171

AN - SCOPUS:0026515407

VL - 148

SP - 267

EP - 271

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 1

ER -