Cloning and characterization of a novel human RNA binding protein gene PNO1.
Present work reported the cloning and characterization of a human novel RNA binding gene Partner of NOB1 (PNO1), with a length of 1637bp and a putative open reading frame of 759 bp, isolated from human kidney. It is composed of seven exons and is localized on chromosome 2p14. Western blot showed that the molecular weight of PNO1 is about 35kDa. RT-PCR results in 16 human tissues indicated that PNO1 is expressed mainly in liver, lung, spleen and kidney, slightly in thymus, testis, ovary, respectively, but not in heart, brain, skeletal muscle, placenta, pancreas, prostate, small intestine, colon and peripheral blood leukocytes. GFP fusion expression in mammalian cells exhibited its localization in the nucleus, especially in nucleoli. Subcellular localization of thirteen GFP fusion PNO1 deletion proteins showed that the region of 92-230 aa is solely responsible for its nucleolar retention, and KH domain alone is not sufficient for nucleolar retention. The PNO1 family shows significant conservation in both eukaryotes and prokaryotes.
Zhou
Guang-Jin
GJ
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, P.R. China.
Zhang
Yue
Y
Wang
Jian
J
Guo
Jin-Hu
JH
Ni
Jun
J
Zhong
Zhao-Ming
ZM
Wang
Li-Qun
LQ
Dang
Yong-Jun
YJ
Dai
Jian-Feng
JF
Yu
Long
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002466:Cell Nucleolus / Q000378:metabolism; D002889:Chromosomes, Human, Pair 2 / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D040461:Gene Components; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008969:Molecular Sequence Data; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; C497714:PNO1 protein, human; D016601:RNA-Binding Proteins; D049452:Green Fluorescent Proteins