Identification of five human novel genes associated with cell proliferation by cell-based screening from an expressed cDNA ORF library.
The development of functional profiling technologies provides opportunity for high-throughput functional genomics studies. We describe a cell-based screening system to identify novel human genes associated with cell proliferation. The method integrates luciferase reporter gene activity, fluorescence stain, automated microscopy and cellular phenotype assays. We successfully used the system to screen 409 novel human genes cloned by our lab and found that 27 genes significantly up-regulated promoter-Renilla luciferase reporter plasmid (pRL) activity. Among them, five genes, TRAF3IP3, ZNF306, ZNF250, SGOL1, and ZNF434, were determined through morphological observation, calcein AM fluorescence stain, MTT assay and cell cycle analysis to be associated with cell proliferation. Furthermore, we showed that the gene TRAF3IP3, which initially was identified to specifically interact with TRAF3, stimulated cell growth by modulating the c-Jun N-terminal kinase (JNK) pathway, and RNAi of TRAF3IP3 confirmed that the effect was physiological and necessary. In summary, we integrated a rapid and efficient system for screening novel growth regulatory genes. Using the new screening system we identified five genes associated with cell proliferation for the first time.
Ma
Xi
X
Chinese National Human Genome Center, Beijing, #3-707 North YongChang Road BDA, Beijing 100176, PR China.
Wang
Xinyu
X
Gao
Xia
X
Wang
Lan
L
Lu
Yang
Y
Gao
Peng
P
Deng
Weiwei
W
Yu
Peng
P
Ma
Jinjing
J
Guo
Jinhai
J
Cheng
Hualing
H
Zhang
Chenying
C
Shi
Taiping
T
Ma
Dalong
D
D002453:Cell Cycle / Q000235:genetics; D049109:Cell Proliferation; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005434:Flow Cytometry; D015723:Gene Library; D015894:Genome, Human; D023281:Genomics; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics; D048008:TNF Receptor-Associated Factor 3 / Q000235:genetics / Q000378:metabolism; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D048008:TNF Receptor-Associated Factor 3; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins; D048031:JNK Mitogen-Activated Protein Kinases