Cloning and characterization of human and mouse SNRK sucrose non-fermenting protein (SNF-1)-related kinases.
We previously isolated, from the earliest population of CD34+ hematopoietic progenitors that form in the aorta of the human embryo, a partial DNA complementary to RNA (cDNA) sequence that was later identified as the human homologue of rat sucrose non-fermenting protein (SNF-1) related kinase (rSNRK), a novel SNF-1-related kinase previously characterized in the rat. In the present study we report the cloning of the complete human SNF-1 related kinase (hSNRK) cDNA and show that the gene spans 39.8 kb at region 3p21 and contains six exons. Recombinant expression of the hSNRK coding sequence in Escherichia coli led to the production of a functional protein kinase of 85 kDa. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of hSNRK expression in fetal CD34+ hematopoietic progenitors revealed its continuous expression throughout human development with higher levels in highly dividing CD34+ CD38+ cells compared to quiescent CD34+ CD38- cells. This observation, together with the expression of hSNRK in numerous human leukemic cell lines, may reflect an implication of hSNRK protein in hematopoietic cell proliferation or differentiation. In the mouse, the SNRK cDNA is 4.6-kb-long and encodes a protein of 748 amino acids with a predicted molecular mass of 81,930 Da. The proteins from human, rat and mouse are strongly conserved and are characterized by the presence of a serine/threonine kinase catalytic domain, a bipartite nuclear targeting signal and an ubiquitin-associated domain. In situ hybridization and RT-PCR analysis of the pattern of mSNRK expression in the mouse reveals that it is temporally and spatially regulated during embryogenesis, and widespread expressed in adult tissues.
Kertesz
Nathalie
N
Howard Hughes Medical Institute and Department of Molecular and Medical Pharmacology, UCLA School of Medicine, Los Angeles, CA 90095-1735, USA.
Samson
Jérôme
J
Debacker
Cécile
C
Wu
Hong
H
Labastie
Marie-Claude
MC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004622:Embryo, Mammalian / Q000201:enzymology / Q000378:metabolism; D005091:Exons; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes / Q000235:genetics; D018922:HL-60 Cells; D006412:Hematopoietic Stem Cells / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D007438:Introns; D019169:Jurkat Cells; D020014:K562 Cells; D008297:Male; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C490993:Snrk protein, mouse; C099821:Snrk protein, rat; C490994:SNRK protein, human; D017346:Protein Serine-Threonine Kinases