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PMID: 14749716 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of the interactions between mammalian PAZ PIWI domain proteins and Dicer.

EMBO reports ·Vol. 5 ·No. 2 ·2004-02-00 ·Pages 189-94

Tahbaz N, Kolb FA, Zhang H, Jaronczyk K, Filipowicz W, Hobman TC

Abstract

PAZ PIWI domain (PPD) proteins, together with the RNA cleavage products of Dicer, form ribonucleoprotein complexes called RNA-induced silencing complexes (RISCs). RISCs mediate gene silencing through targeted messenger RNA cleavage and translational suppression. The PAZ domains of PPD and Dicer proteins were originally thought to mediate binding between PPD proteins and Dicer, although no evidence exists to support this theory. Here we show that PAZ domains are not required for PPD protein-Dicer interactions. Rather, a subregion of the PIWI domain in PPD proteins, the PIWI-box, binds directly to the Dicer RNase III domain. Stable binding between PPD proteins and Dicer was dependent on the activity of Hsp90. Unexpectedly, binding of PPD proteins to Dicer inhibits the RNase activity of this enzyme in vitro. Lastly, we show that PPD proteins and Dicer are present in soluble and membrane-associated fractions, indicating that interactions between these two types of proteins may occur in multiple compartments.

MeSH Terms
Animals Argonaute Proteins Binding Sites Eukaryotic Initiation Factor-2 Glutathione Transferase/analysis,genetics HSP90 Heat-Shock Proteins/metabolism Humans Peptide Initiation Factors/chemistry,metabolism Protein Structure, Tertiary/genetics Proteins/chemistry,genetics,metabolism RNA Interference RNA, Double-Stranded/metabolism RNA-Induced Silencing Complex/genetics,metabolism Rats Ribonuclease III/chemistry,genetics,metabolism Two-Hybrid System Techniques
Chemicals
AGO2 protein, human Argonaute Proteins Eukaryotic Initiation Factor-2 HSP90 Heat-Shock Proteins PIWIL1 protein, human Peptide Initiation Factors Proteins RNA, Double-Stranded RNA-Induced Silencing Complex Glutathione Transferase Ribonuclease III
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Tahbaz Nasser
Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada.
Kolb Fabrice A
Zhang Haidi
Jaronczyk Katarzyna
Filipowicz Witold
Hobman Tom C
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Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-221X
DOI
10.1038/sj.embor.7400070
Published
2004-02-00
Epub
2004-00-16
Pages
189-94
Language
English
Region
England
NLM ID
100963049
PMCID
PMC1298981
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.embor.7400070 PMC Full Text View journal details
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