Molecular cloning of cDNAs for the human granulocyte colony-stimulating
factor receptor from HL-60 and mapping of the gene to chromosome region
1p32-34
DJ Tweardy, K Anderson, LA Cannizzaro, RA Steinman, CM Croce and K Huebner
Department of Medicine, University of Pittsburgh School of Medicine, PA.
Early studies examining the effects of purified or recombinant granulocyte
colony-stimulating factor (G-CSF) on human leukemia cell lines demonstrated
that some cell lines, such as HL-60, could be induced to differentiate in
response to G-CSF. In two recent studies reporting the cloning of the human
G-CSF receptor (hGCSFR), four classes of receptor cDNA were identified and,
surprisingly, the message for this receptor was reportedly expressed by
HL-60 at either very low levels or not at all. Using a mouse G-CSF receptor
probe, we cloned and sequenced a cDNA for hGCSFR from an HL-60 cDNA library
in plasmid and used it to identify 31 additional clones from an HL-60 cDNA
library in phage. Polymerase chain reaction analysis of the 31 phage clones
established that 29 were derived from class I hGCSFR mRNA, one was derived
from class III mRNA, and one was derived from class IV mRNA. In addition,
the hGCSFR gene was chromosomally localized by Southern blot analysis of
its segregation pattern in a panel of rodent-human hybrid DNAs using the
radiolabeled cDNA probe. The hGCSFR locus was present in hybrids retaining
the distal short arm of human chromosome 1 and absent in hybrids that did
not retain this region. Chromosomal in situ hybridization refined the
localization of the hGCSFR gene to region 1p32-p34. The combination of
hybrid DNA analysis and in situ hybridization places the hGCSFR gene
telomeric to the CSF1, JUN, and TCL-5 loci.
Volume 79,
Issue 5,
pp. 1148-1154,
03/01/1992
Copyright © 1992 by The American Society of Hematology
