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Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
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hPop5, a protein subunit of the human RNase MRP and RNase P endoribonucleases

H van Eenennaam1, D Lugtenberg, J H Vogelzangs

  • 1Department of Biochemistry, University of Nijmegen, P. O. Box 9101, NL-6500 HB Nijmegen, The Netherlands.

The Journal of Biological Chemistry
|June 20, 2001
PubMed

Insights

Researchers identified the conserved Pop5 protein subunit in human, rat, mouse, cow, and Drosophila. This protein is crucial for RNase MRP and RNase P functions in precursor RNA processing.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • RNase MRP and RNase P are ribonucleoprotein endoribonucleases.
  • RNase MRP processes precursor-rRNA, while RNase P processes pre-tRNA.
  • Both complexes share RNA and protein components with similar structures.

Purpose of the Study:

  • To identify and characterize homologues of the Pop5 protein subunit.
  • To investigate the association and localization of human Pop5 (hPop5) within RNase MRP and RNase P complexes.
  • To determine the role of hPop5 in the catalytic activity and complex formation of RNase P.

Main Methods:

  • Homologue identification across species.
  • cDNA cloning and protein expression.
  • Antibody generation and western blotting.
  • Immunofluorescence microscopy for protein localization.
  • Partial purification of RNase P and co-immunoprecipitation assays.

Main Results:

  • Human, rat, mouse, cow, and Drosophila homologues of Pop5p were identified.
  • Recombinant hPop5 antibodies detected a 19-kDa polypeptide in HeLa cells, associated with both RNase MRP and RNase P.
  • hPop5 localizes to the nucleus and nucleolus.
  • hPop5 is associated with catalytically active RNase P.
  • The C-terminal tail of hPop5 is not essential for complex formation or RNase P activity.

Conclusions:

  • The Pop5 protein is an evolutionarily conserved subunit of RNase MRP and RNase P.
  • hPop5 is a nuclear and nucleolar protein associated with both RNase complexes.
  • hPop5 plays a role in RNase P function and complex integrity.

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