[Preliminary structural and functional study on a novel gene HSPCSET].
OBJECTIVE
To characterize the structural and the functional feature of a novel gene HSPCSET isolated from human CD34+ hematopoietic stem/progenitor cells (HS/PCs).


METHODS
Bioinformatic technology was used to identify the structural features of the HSPCSET protein and perform the multiple sequence alignment. Yeast-two-hybrid system was used to identify the proteins interacting with the HSPCSET protein. After sequencing, we selected out the positive clones which had clear functions, and carried out beta-gal experiment and GST pull down assay to confirm the results. The cellular location of the HSPCSET was checked by immunofluorescence assay.


RESULTS
The HSPCSET protein belongs to a SET domain family, which is evolutionarily conserved across species. It implied that HSPCSET may have biologically important function. Using yeast-two-hybrid system, we showed that the protein sequence with SET domain might bind to 13 proteins, which involved in signaling transduction, transcriptional regulation, apoptosis, tumorigenesis, development, etc. And 4 proteins (GADD34, SIVA, DNAJ and PHF1) were confirmed by one-on-one back of the hybrid experiment, beta-gal test and GST pull down assay. When GADD34 and HSPCSET were co-transfected, they co-localized in the nucleus, suggesting a strong interaction.


CONCLUSIONS
The novel gene HSPCSET is likely to have biologically important function. This study provides the basis for further studies of its function in hematopoiesis and tumorigenesis.
Wei
Ju
J
Department of Hematology, Shanghai First People's Hospital, Shanghai, 200080 PR China.
Sun
Xiao-jian
XJ
Wu
Xin-yan
XY
Chen
Sai-juan
SJ
Chen
Zhu
Z
Wang
Chun
C
Huang
Qiu-hua
QH
D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D019295:Computational Biology; D017124:Conserved Sequence; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D054645:Protein Phosphatase 1; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
D000943:Antigens, Differentiation; D018797:Cell Cycle Proteins; D011506:Proteins; C087519:PPP1R15A protein, human; D054645:Protein Phosphatase 1