Identification and characterization of a novel cytoplasm protein ICF45 that is involved in cell cycle regulation.
A novel cytoplasm protein, interphase cytoplasm foci protein 45 kDa (ICF45), was identified by screening the cDNA expression library for HeLa cells with serum from an autoimmune patient. The complete cDNA sequence of ICF45 was determined to be 1.32 kb and to encode 298 amino acids with an apparent molecular mass of 45 kDa. The ICF45 transcripts were detected in different tissues and were relatively rich in human liver and lung tissues but scarce in brain tissue. Immunofluorescence with anti-ICF45-specific antibodies demonstrated that ICF45 is strongly expressed in interphase and cannot be seen in mitosis. The subcellular localization of ICF45 and fusion proteins GFP-ICF45, ICF45-GFP, and HA-ICF45 showed ICF45 centralized into 1-2 dots in the cytoplasm and always near the nuclear membrane. The staining foci of ICF45 appeared to be slightly larger than centrosomes and in some cases were found to colocalize with centrosomes. After effectively silencing the ICF45 by RNAi, the growth and proliferation of the cells were significantly inhibited, and p53 was detected to be up-regulated. The silencing of ICF45 also resulted in an appearance of polycentrosome and multinuclear cells, which finally went to apoptosis. Our results suggest that ICF45 is a highly conserved novel protein, which is expressed in a cell cycle-dependent manner and seemed to be involved in cell cycle progression and cell proliferation.
The Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Science, Beijing Normal University, Beijing 100875, China.
Deliang
D
Guo
Kun
K
Hu
Ying
Y
Lei
Yongchao
Y
Wang
Tianlin
T
Ma
Dacheng
D
He
D000595:Amino Acid Sequence; D017209:Apoptosis; D001483:Base Sequence; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002453:Cell Cycle; D049109:Cell Proliferation; D018385:Centrosome / Q000648:ultrastructure; D003062:Codon; D017124:Conserved Sequence; D003593:Cytoplasm / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D015723:Gene Library; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008938:Mitosis; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015854:Up-Regulation
D003062:Codon; D018076:DNA, Complementary; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53; D019814:Myristic Acid; D049452:Green Fluorescent Proteins