Abstract
A yeast two-hybrid system was used to analyze interactions among the protein subunits of human nuclear RNase P themselves and with other interacting partners encoded in a HeLa cell cDNA library. Subunits hpop1, Rpp21, Rpp29, Rpp30, Rpp38, and Rpp40 are involved in extensive, but weak, protein-protein interactions in the holoenzyme complex. Rpp14, Rpp20, and Rpp30 were found to have strong interactions with proteins encoded in the cDNA library. The small heat shock protein 27, which interacts with Rpp20 in the two-hybrid assay, binds to Rpp20 during affinity chromatography and can be found to be associated with, and enhances the activity of, highly purified RNase P. RNase P activity in HeLa cell nuclei also increases under the stress of heat shock.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Autoantigens / chemistry
-
Autoantigens / metabolism
-
Binding Sites
-
Cell Nucleus / enzymology
-
Endoribonucleases / chemistry*
-
Endoribonucleases / genetics
-
Endoribonucleases / metabolism*
-
Enzyme Activation
-
Gene Library
-
HSP27 Heat-Shock Proteins
-
HeLa Cells
-
Heat-Shock Proteins / metabolism
-
Humans
-
Kinetics
-
Models, Molecular
-
Molecular Chaperones
-
Neoplasm Proteins / metabolism
-
Protein Subunits
-
RNA, Catalytic / chemistry*
-
RNA, Catalytic / genetics
-
RNA, Catalytic / metabolism*
-
Recombinant Proteins / chemistry
-
Recombinant Proteins / metabolism
-
Ribonuclease P
-
Thermodynamics
Substances
-
Autoantigens
-
HSP27 Heat-Shock Proteins
-
HSPB1 protein, human
-
Heat-Shock Proteins
-
Molecular Chaperones
-
Neoplasm Proteins
-
POP7 protein, human
-
Protein Subunits
-
RNA, Catalytic
-
Recombinant Proteins
-
Endoribonucleases
-
RPP14 protein, human
-
Ribonuclease P