Human dicer: purification, properties, and interaction with PAZ PIWI domain proteins.
Dicer is a multidomain ribonuclease that processes double-stranded RNAs (dsRNAs) to 21-nt small interfering RNAs (siRNAs) during RNA interference and excises microRNAs (miRNAs) from precursor hairpins. PAZ and PIWI domain (PPD) proteins, also involved in RNAi and miRNA function, are the best-characterized proteins known to interact with Dicer. PPD proteins are the core constituents of effector complexes, RISCs and miRNPs, mediating siRNA and miRNA function. In this chapter we describe overexpression and purification of recombinant human Dicer, its biochemical properties, and mapping of domains responsible for Dicer-PPD protein interactions.
Kolb
Fabrice A
FA
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
Zhang
Haidi
H
Jaronczyk
Kasia
K
Tahbaz
Nasser
N
Hobman
Tom C
TC
Filipowicz
Witold
W
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D043244:Ribonuclease III / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D011506:Proteins; D011994:Recombinant Proteins; D043244:Ribonuclease III