Crystal structure of the primary piRNA biogenesis factor Zucchini reveals similarity to the bacterial PLD endonuclease Nuc
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Crystal structure of the primary piRNA biogenesis factor Zucchini reveals similarity to the bacterial PLD endonuclease Nuc. / Voigt, Franka; Reuter, Michael; Kasaruho, Anisa; Schulz, Eike C; Pillai, Ramesh S; Barabas, Orsolya.
In: RNA, Vol. 18, No. 12, 12.2012, p. 2128-34.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Crystal structure of the primary piRNA biogenesis factor Zucchini reveals similarity to the bacterial PLD endonuclease Nuc
AU - Voigt, Franka
AU - Reuter, Michael
AU - Kasaruho, Anisa
AU - Schulz, Eike C
AU - Pillai, Ramesh S
AU - Barabas, Orsolya
PY - 2012/12
Y1 - 2012/12
N2 - 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.
AB - 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.
KW - Amino Acid Sequence
KW - Animals
KW - Bacterial Proteins/chemistry
KW - Catalytic Domain
KW - Crystallography, X-Ray
KW - Drosophila Proteins/chemistry
KW - Drosophila melanogaster/enzymology
KW - Endoribonucleases/chemistry
KW - Mice
KW - Mitochondrial Proteins/chemistry
KW - Models, Molecular
KW - Molecular Sequence Data
KW - Phospholipase D/chemistry
KW - Protein Multimerization
KW - RNA, Small Interfering/biosynthesis
KW - Sequence Homology, Amino Acid
KW - Static Electricity
U2 - 10.1261/rna.034967.112
DO - 10.1261/rna.034967.112
M3 - SCORING: Journal article
C2 - 23086923
VL - 18
SP - 2128
EP - 2134
JO - RNA
JF - RNA
SN - 1355-8382
IS - 12
ER -
