Sequence analysis and functional studies of interleukin-3 receptor α subunit-encoding cDNAs amplified from KG-1 leukemic cells and normal human marrow

Aaron P. Rapoport; John F. DiPersio

doi:10.1016/0378-1119(93)90030-7

Sequence analysis and functional studies of interleukin-3 receptor α subunit-encoding cDNAs amplified from KG-1 leukemic cells and normal human marrow

Aaron P. Rapoport, John F. DiPersio

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Two partial cDNAs encoding the human interleukin-3 receptor a subunit (IL-3Rα) were cloned from KG-1 leukemic cells using the polymerase chain reaction. Sequence analysis of these cDNAs predicted two single-amino-acid (aa) changes as compared with the published TF-1 leukemic cell IL-3Rα sequence [Kitamura et al., Cell 66 (1991) 1165-1174]. These changes were confirmed by sequence analysis of a second set of independently derived cDNAs. Identical aa changes were found in the IL-3Rα encoded by cDNAs cloned from normal CD34⁺ human marrow cells. Ligation of the partial cDNAs derived from KG-1 cells resulted in a full-length functional IL-3Rα cDNA clone. Deletion of the extracellular 'LSXWS' consensus sequence resulted in complete loss of detectable [¹²⁵I]IL-3 binding when this mutant receptor was co-expressed in COS-7 cells with the β subunit of the IL-3 receptor.

Original language	English
Pages (from-to)	333-337
Number of pages	5
Journal	Gene
Volume	137
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(93)90030-7
State	Published - Dec 31 1993

Keywords

Hematopoietic growth factors
amino acid polymorphism
cDNA cloning
cytokine receptor
mutational analysis
polymerase chain reaction

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10.1016/0378-1119(93)90030-7

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@article{c05199745872407ca872a25e6420d917,

title = "Sequence analysis and functional studies of interleukin-3 receptor α subunit-encoding cDNAs amplified from KG-1 leukemic cells and normal human marrow",

abstract = "Two partial cDNAs encoding the human interleukin-3 receptor a subunit (IL-3Rα) were cloned from KG-1 leukemic cells using the polymerase chain reaction. Sequence analysis of these cDNAs predicted two single-amino-acid (aa) changes as compared with the published TF-1 leukemic cell IL-3Rα sequence [Kitamura et al., Cell 66 (1991) 1165-1174]. These changes were confirmed by sequence analysis of a second set of independently derived cDNAs. Identical aa changes were found in the IL-3Rα encoded by cDNAs cloned from normal CD34+ human marrow cells. Ligation of the partial cDNAs derived from KG-1 cells resulted in a full-length functional IL-3Rα cDNA clone. Deletion of the extracellular 'LSXWS' consensus sequence resulted in complete loss of detectable [125I]IL-3 binding when this mutant receptor was co-expressed in COS-7 cells with the β subunit of the IL-3 receptor.",

keywords = "Hematopoietic growth factors, amino acid polymorphism, cDNA cloning, cytokine receptor, mutational analysis, polymerase chain reaction",

author = "Rapoport, {Aaron P.} and DiPersio, {John F.}",

note = "Funding Information: The authors gratefully acknowledge the gift of pKH97 from Dr. A. Miyajima (DNAX Research Institute of Molecular and Cellular Biology, Palo Alto, CA, USA). This work was supported by Public Health Service Funding Information: Training Grant No. 5T-32-HL-07152-18 (APR), and grants BE-98B and JFRA-285 from the American Cancer Society (JFD). The authors thank Carol Weed for help in preparing the manuscript.",

year = "1993",

month = dec,

day = "31",

doi = "10.1016/0378-1119(93)90030-7",

language = "English",

volume = "137",

pages = "333--337",

journal = "Gene",

issn = "0378-1119",

number = "2",

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T1 - Sequence analysis and functional studies of interleukin-3 receptor α subunit-encoding cDNAs amplified from KG-1 leukemic cells and normal human marrow

AU - Rapoport, Aaron P.

AU - DiPersio, John F.

N1 - Funding Information: The authors gratefully acknowledge the gift of pKH97 from Dr. A. Miyajima (DNAX Research Institute of Molecular and Cellular Biology, Palo Alto, CA, USA). This work was supported by Public Health Service Funding Information: Training Grant No. 5T-32-HL-07152-18 (APR), and grants BE-98B and JFRA-285 from the American Cancer Society (JFD). The authors thank Carol Weed for help in preparing the manuscript.

PY - 1993/12/31

Y1 - 1993/12/31

N2 - Two partial cDNAs encoding the human interleukin-3 receptor a subunit (IL-3Rα) were cloned from KG-1 leukemic cells using the polymerase chain reaction. Sequence analysis of these cDNAs predicted two single-amino-acid (aa) changes as compared with the published TF-1 leukemic cell IL-3Rα sequence [Kitamura et al., Cell 66 (1991) 1165-1174]. These changes were confirmed by sequence analysis of a second set of independently derived cDNAs. Identical aa changes were found in the IL-3Rα encoded by cDNAs cloned from normal CD34+ human marrow cells. Ligation of the partial cDNAs derived from KG-1 cells resulted in a full-length functional IL-3Rα cDNA clone. Deletion of the extracellular 'LSXWS' consensus sequence resulted in complete loss of detectable [125I]IL-3 binding when this mutant receptor was co-expressed in COS-7 cells with the β subunit of the IL-3 receptor.

AB - Two partial cDNAs encoding the human interleukin-3 receptor a subunit (IL-3Rα) were cloned from KG-1 leukemic cells using the polymerase chain reaction. Sequence analysis of these cDNAs predicted two single-amino-acid (aa) changes as compared with the published TF-1 leukemic cell IL-3Rα sequence [Kitamura et al., Cell 66 (1991) 1165-1174]. These changes were confirmed by sequence analysis of a second set of independently derived cDNAs. Identical aa changes were found in the IL-3Rα encoded by cDNAs cloned from normal CD34+ human marrow cells. Ligation of the partial cDNAs derived from KG-1 cells resulted in a full-length functional IL-3Rα cDNA clone. Deletion of the extracellular 'LSXWS' consensus sequence resulted in complete loss of detectable [125I]IL-3 binding when this mutant receptor was co-expressed in COS-7 cells with the β subunit of the IL-3 receptor.

KW - Hematopoietic growth factors

KW - amino acid polymorphism

KW - cDNA cloning

KW - cytokine receptor

KW - mutational analysis

KW - polymerase chain reaction

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U2 - 10.1016/0378-1119(93)90030-7

DO - 10.1016/0378-1119(93)90030-7

M3 - Article

C2 - 8299967

AN - SCOPUS:0027892031

SN - 0378-1119

VL - 137

SP - 333

EP - 337

JO - Gene

JF - Gene

IS - 2

ER -

Sequence analysis and functional studies of interleukin-3 receptor α subunit-encoding cDNAs amplified from KG-1 leukemic cells and normal human marrow

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