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Molecular cloning of a novel receptor tyrosine kinase gene, STK, derived
from enriched hematopoietic stem cells
A Iwama, K Okano, T Sudo, Y Matsuda and T Suda
Department of Cell Differentiation, Kumamoto University School of Medicine,
Japan.
To identify the novel receptor tyrosine kinases (RTKs) critical to the
proliferation of hematopoietic stem cells, we performed polymerase chain
reaction-based cloning from highly purified murine hematopoietic stem
cells. Lineage marker-negative, c-KIT-positive, and Ly6A/E- or Sca-
1-positive (Lin-c-KIT+Sca-1+) cells were sorted by a fluorescence-
activated cell sorter. Two sets of degenerate oligonucleotide primers were
directed to the conserved sequences of the catalytic domain, and were used
to amplify cDNAs that encode protein tyrosine kinases (PTKs). One hundred
cDNA clones were sequenced and 8 RTKs were identified, as well as 12
non-RTKs and 2 serine/threonine kinases. Sixteen cDNAs were identical to
the known kinase genes (PKC beta, JAK-1, JAK-2, TYK-2, HCK, FGR, FYN, BLK,
c-FES, FER, c-ABL, c-KIT, FLK-1, FLK-2, IGF1R, and ECK). Six novel cDNA
sequences (stk series) were identified. However, three of them turned out
to be BPK, RYK, and TEK. The remaining three showed high homology to S6
kinase II, JAK-2, and v-SEA/c-MET, respectively. Characterization of
full-length cDNA sequence of the v- SEA/cMET-related gene showed that this
was a novel RTK gene and we named this gene STK (stem cell-derived tyrosine
kinase). We identified two distinct forms of STK cDNA; the short one
encoded a putative truncated protein that lacked most of the extracellular
domain. STK was expressed at various stages of hematopoietic cells,
including stem cells, but we could not detect any apparent expression in
other adult tissues. The expression of the truncated form of mRNA was more
predominant than that of the complete form. STK was assigned by fluorescent
in situ hybridization to the R-positive F1 band of chromosome 9, the same
region to which hepatic growth factor-like protein has been assigned.
Characterization of these PTKs, including STK, will be helpful to elucidate
the molecular mechanism of the growth regulation of hematopoietic stem
cells.
Volume 83,
Issue 11,
pp. 3160-3169,
06/01/1994
Copyright © 1994 by The American Society of Hematology
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