Purification and characterization of Rpp25, an RNA-binding protein subunit of human ribonuclease P.
In HeLa cells, ribonuclease P (RNase P), the tRNA processing enzyme consists of an RNA subunit (H1 RNA) associated with at least nine protein subunits, Rpp14, Rpp20, Rpp21, Rpp29 (hPop4), Rpp30, Rpp38, Rpp40, hPop1, and hPop5 (18.8 kDa). We report here the cloning and immuno-biochemical analysis of Rpp25, another protein subunit of RNase P. Polyclonal rabbit antibodies raised against recombinant Rpp25 recognize their corresponding antigens in RNase P-containing fractions purified from HeLa cells, and they also precipitate active holoenzyme. Furthermore, this protein has general RNA binding properties.
Guerrier-Takada
Cecilia
C
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520, USA.
Eder
Paul S
PS
Gopalan
Venkat
V
Altman
Sidney
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D024202:Electrophoretic Mobility Shift Assay; D004722:Endoribonucleases / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D021122:Protein Subunits; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification*; D043262:Ribonuclease P; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays
D017931:DNA Primers; D021122:Protein Subunits; D016337:RNA, Catalytic; D016601:RNA-Binding Proteins; C455399:RPP25 protein, human; D004722:Endoribonucleases; C468368:RPP14 protein, human; D043262:Ribonuclease P