ABSTRACT
Objective To screen and identify proteins interacting with hematopoietic stem/progenitor cell differentiation-related gene HSPC016, and to explore the molecular mechanism involved in the regulation by HSPC016 on the aggregative behavior of dermal papilla cells. Methods By using yeast two-hybridization,HSPC016 gene was sub-cloned into pGBKT7 to construct the bait plasmid (named as pGBKT7-HSPC016) in yeast AH109. The cDNA yeast expression library of human dermal papillae cells in yeast Y187 was screened with the bait plasmid and the proteins interacting with HSPC016 were identified. Yeast two-hybridization retransformation experiment was conducted to exclude the false positive clones and verify the interactions, then,the positive clones were sequenced and analyzed by using bioinformatic methods. Results The bait plasmid pGBKT7-HSPC016 was constructed successfully and there was no self-activation in or toxicity against yeast AH 109. Four proteins,including forkhead family of transcription factors (FOXO 1 ), mitogen-activated protein kinase 11 (MAPK 11 ), phosphoinositide-3-kinase (PIK3R3) and liver X receptor were screened and identified. Bioinformatic analysis revealed that these proteins had close relationship with intracellular energy metabolism and translational regulation. Conclusions HSPC016 may regulate the aggregative behavior of DPCs by regulating the levels of intracellular reactive oxygen species (ROS) and interacting with signaling molecules involved in intracellular energy metabolism and translational regulation.
ABSTRACT
Objective To clone the full length cDNA sequence of HSPC016 gene, an aggregative growth related gene in dermal papilla cells (DPC). Analyze its characteristics and predict its biological function in the phase of growth and differentiation for DPC. Methods Rapid amplification of cDNA ends (RACE) technology was used for full length cDNA amplification. Bioinformtic methods were used to analyze the chromosome location, protein sequence, domain and possible biological function of the gene. Results Two isoforms of HSPC016 gene were obtained from DPC. They were 400bp and 493bp, respectively. The gene was mapped on chromosome 3 q21 31, and was conservative on evolution. HSPC016 protein had 64aa, belonged to PD053992 protein family; its functional domain was homologous to T2FA gene. Conclusion HSPC016 was a transcriptional modulatory gene. Its protein product may act as a subunit of a transcriptional complex and play a role on DPC growth and differentiation through modulating other genes' transcription within nucleus