The structural biochemistry of Zucchini implicates it as a nuclease in piRNA biogenesis

Ipsaro, J. J., Haase, A. D., Knott, S. R., Joshua-Tor, L., Hannon, G. J. (2012) The structural biochemistry of Zucchini implicates it as a nuclease in piRNA biogenesis. Nature, 491 (7423). pp. 279-282. ISSN 00280836 (ISSN) (Public Dataset)

Abstract

PIWI-family proteins and their associated small RNAs (piRNAs) act in an evolutionarily conserved innate immune mechanism to provide essential protection for germ-cell genomes against the activity of mobile genetic elements. piRNA populations comprise a molecular definition of transposons, which permits them to distinguish transposons from host genes and selectively silence them. piRNAs can be generated in two distinct ways, forming either primary or secondary piRNAs. Primary piRNAs come from discrete genomic loci, termed piRNA clusters, and seem to be derived from long, single-stranded precursors. The biogenesis of primary piRNAs involves at least two nucleolytic steps. An unknown enzyme cleaves piRNA cluster transcripts to generate monophosphorylated piRNA 5′ ends. piRNA 3′ ends are probably formed by exonucleolytic trimming, after a piRNA precursor is loaded into its PIWI partner. Secondary piRNAs arise during the adaptive 'ping-pong' cycle, with their 5′ termini being formed by the activity of PIWIs themselves. A number of proteins have been implicated genetically in primary piRNA biogenesis. One of these, Drosophila melanogaster Zucchini, is a member of the phospholipase-D family of phosphodiesterases, which includes both phospholipases and nucleases. Here we produced a dimeric, soluble fragment of the mouse Zucchini homologue (mZuc; also known as PLD6) and show that it possesses single-strand-specific nuclease activity. A crystal structure of mZuc at 1.75Å resolution indicates greater architectural similarity to phospholipase-D family nucleases than to phospholipases. Together, our data suggest that the Zucchini proteins act in primary piRNA biogenesis as nucleases, perhaps generating the 5′ ends of primary piRNAs. © 2012 Macmillan Publishers Limited. All rights reserved.

Item Type:	Paper
Uncontrolled Keywords:	Drosophila protein nuclease phospholipase D Piwi interacting RNA unclassified drug Zucchini protein biochemistry cell organelle enzyme activity fly genome immune response protein RNA vegetable animal cell article controlled study crystal structure nonhuman priority journal protein function protein structure RNA synthesis Cucurbita pepo var. melopepo Drosophila melanogaster
Subjects:	bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > piRNA
CSHL Authors:	Haase, Astrid D. Hannon, Gregory J. Ipsaro, Jonathan J. Joshua-Tor, Leemor Knott, Simon R.V.
Communities:	CSHL Cancer Center Program > Cancer Genetics CSHL Cancer Center Program > Gene Regulation and Cell Proliferation CSHL Cancer Center Shared Resources > DNA Sequencing Service CSHL Cancer Center Shared Resources > Mass Spectrometry Service CSHL Cancer Center Shared Resources > Proteomics Service CSHL Post Doctoral Fellows CSHL labs > Hannon lab CSHL labs > Joshua-Tor lab
Depositing User:	Matt Covey
Date:	2012
Date Deposited:	09 Jan 2013 16:11
Last Modified:	05 Sep 2017 15:58
PMCID:	PMC3493678
Related URLs:	Publisher
Dataset ID:	PDB: 4GGJ PDB: 4GGK
URI:	https://repository.cshl.edu/id/eprint/26416

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