Molecular cloning of murine STAP-1, the stem-cell-specific adaptor protein containing PH and SH2 domains

Biochem Biophys Res Commun. 2000 Feb 24;268(3):697-703. doi: 10.1006/bbrc.2000.2223.


To identify the novel substrate of c-kit which is important for hematopoietic stem cell self-renewal or differentiation, CD34-low/negative, Sca-1-positive, c-kit-positive, and lineage marker-negative (CD34(low/-)Sca-1(+)c-kit(+)Lin(-)) cells were sorted by a fluorescence-activated cell sorter from mouse bone marrow cells and a yeast two-hybrid cDNA library was constructed. By screening with c-kit as bait, we cloned a novel cDNA, designed STAP-1, encoding an adaptor protein with a Pleckstrin homology domain, the Src homology 2 (SH2) domain, and a number of tyrosine phosphorylation sites. RT-PCR analysis revealed that STAP-1 expression is restricted in the bone marrow cell fraction expressing c-kit. The highest expression was observed in the CD34(low/-)Sca-1(+)c-kit(+)Lin(-) stem cell-enriched fraction. The murine myeloid cell line, M1, expressed a high level of STAP-1. However, the expression was strongly repressed in response to leukemia inhibitory factor (LIF) which induced monocytic differentiation of M1 cells, suggesting that STAP-1 is associated with the undifferentiated cell type. A two-hybrid assay indicated that STAP-1 bound not only to c-kit but also to c-fms but not to JAK2 or Pyk2. In 293 cells, STAP-1 was tyrosine-phosphorylated by activated c-kit. An in vitro binding assay suggested that the STAP-1 SH2 domain interacted with several tyrosine-phosphorylated proteins including c-kit and STAT5. These suggest that STAP-1 functions as an adaptor molecule downstream of c-kit in hematopoietic stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, CD34 / metabolism
  • Antigens, Ly / metabolism
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics
  • Gene Expression
  • Hematopoietic Stem Cells / immunology
  • Hematopoietic Stem Cells / metabolism*
  • Membrane Proteins / metabolism
  • Mice
  • Molecular Sequence Data
  • Phosphorylation
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-kit / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Homology, Amino Acid
  • src Homology Domains


  • Antigens, CD34
  • Antigens, Ly
  • DNA Primers
  • Ly6a protein, mouse
  • Membrane Proteins
  • RNA, Messenger
  • Proto-Oncogene Proteins c-kit

Associated data

  • GENBANK/AB036058