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PMID: 23990920 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Human RNase H1 is associated with protein P32 and is involved in mitochondrial pre-rRNA processing.

PloS one ·Vol. 8 ·No. 8 ·2013-00-00 ·Pages e71006

Wu H, Sun H, Liang X, Lima WF, Crooke ST

Abstract

Mammalian RNase H1 has been implicated in mitochondrial DNA replication and RNA processing and is required for embryonic development. We identified the mitochondrial protein P32 that binds specifically to human RNase H1, but not human RNase H2. P32 binds human RNase H1 via the hybrid-binding domain of the enzyme at an approximately 1∶1 ratio. P32 enhanced the cleavage activity of RNase H1 by reducing the affinity of the enzyme for the heteroduplex substrate and enhancing turnover, but had no effect on the cleavage pattern. RNase H1 and P32 were partially co-localized in mitochondria and reduction of P32 or RNase H1 levels resulted in accumulation of mitochondrial pre ribosomal RNA [12S/16S] in HeLa cells. P32 also co-immunoprecipitated with MRPP1, a mitochondrial RNase P protein required for mitochondrial pre-rRNA processing. The P32-RNase H1 complex was shown to physically interact with mitochondrial DNA and pre-rRNA. These results expand the potential roles for RNase H1 to include assuring proper transcription and processing of guanosine-cytosine rich pre-ribosomal RNA in mitochondria. Further, the results identify P32 as a member of the 'RNase H1 degradosome' and the key P32 enhances the enzymatic efficiency of human RNase H1.

MeSH Terms
Carrier Proteins Cell Line Gene Expression Regulation, Enzymologic HEK293 Cells HeLa Cells Humans Hydrolysis Mass Spectrometry Microscopy, Fluorescence Mitochondria/metabolism Mitochondrial Proteins/metabolism RNA Interference RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA, Ribosomal/metabolism Ribonuclease H/metabolism
Chemicals
C1QBP protein, human Carrier Proteins Mitochondrial Proteins RNA Precursors RNA, Ribosomal Ribonuclease H ribonuclease HI
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wu Hongjiang
Department of Core Antisense Research, Isis Pharmaceuticals, Inc., Carlsbad, California, United States of America.
Sun Hong
Liang Xuehai
Lima Walt F
Crooke Stanley T
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Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0071006
Published
2013-00-00
Epub
2013-00-22
Pages
e71006
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3750045
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0071006 PMC Full Text View journal details
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