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  • Crystal structure of the primary piRNA biogenesis factor Zucchini reveals similarity to the bacterial PLD endonuclease Nuc.

Crystal structure of the primary piRNA biogenesis factor Zucchini reveals similarity to the bacterial PLD endonuclease Nuc.

RNA (New York, N.Y.) (2012-10-23)
Franka Voigt, Michael Reuter, Anisa Kasaruho, Eike C Schulz, Ramesh S Pillai, Orsolya Barabas
ABSTRACT

Piwi-interacting RNAs (piRNAs) are a gonad-specific class of small RNAs that associate with the Piwi clade of Argonaute proteins and play a key role in transposon silencing in animals. Since biogenesis of piRNAs is independent of the double-stranded RNA-processing enzyme Dicer, an alternative nuclease that can process single-stranded RNA transcripts has been long sought. A Phospholipase D-like protein, Zucchini, that is essential for piRNA processing has been proposed to be a nuclease acting in piRNA biogenesis. Here we describe the crystal structure of Zucchini from Drosophila melanogaster and show that it is very similar to the bacterial endonuclease, Nuc. The structure also reveals that homodimerization induces major conformational changes assembling the active site. The active site is situated on the dimer interface at the bottom of a narrow groove that can likely accommodate single-stranded nucleic acid substrates. Furthermore, biophysical analysis identifies protein segments essential for dimerization and provides insights into regulation of Zucchini's activity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phospholipase D from cabbage, Type IV, lyophilized powder, ≥100 units/mg solid
Pricing and availability is not currently available.
Sigma-Aldrich
Phospholipase D from Arachis hypogaea (peanut), Type II, lyophilized powder, ≥60 units/mg protein
Pricing and availability is not currently available.
Sigma-Aldrich
Phospholipase D from Streptomyces sp., Type VII, lyophilized powder, ≥150 units/mg solid
Pricing and availability is not currently available.
Sigma-Aldrich
Phospholipase D from Streptomyces chromofuscus, ≥50,000 units/mL, buffered aqueous glycerol solution
Pricing and availability is not currently available.