Functional involvement of Tudor and dPRMT5 in the piRNA processing pathway in Drosophila germlines.
In Drosophila, the PIWI proteins, Aubergine (Aub), AGO3, and Piwi are expressed in germlines and function in silencing transposons by associating with PIWI-interacting RNAs (piRNAs). Recent studies show that PIWI proteins contain symmetric dimethyl-arginines (sDMAs) and that dPRMT5/Capsuleen/DART5 is the modifying enzyme. Here, we show that Tudor (Tud), one of Tud domain-containing proteins, associates with Aub and AGO3, specifically through their sDMA modifications and that these three proteins form heteromeric complexes. piRNA precursor-like molecules are detected in these complexes. The expression levels of Aub and AGO3, along with their degree of sDMA modification, were not changed by tud mutations. However, the population of transposon-derived piRNAs associated with Aub and AGO3 was altered by tud mutations, whereas the total amounts of small RNAs on Aub and AGO3 was increased. Loss of dprmt5 did not change the stability of Aub, but impaired its association with Tud and lowered piRNA association with Aub. Thus, in germline cells, piRNAs are quality-controlled by dPRMT5 that modifies PIWI proteins, in tight association with Tud.
Nishida
Kazumichi M
KM
Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
Okada
Tomoko N
TN
Kawamura
Takeshi
T
Mituyama
Toutai
T
Kawamura
Yoshinori
Y
Inagaki
Sachi
S
Huang
Haidong
H
Chen
Dahua
D
Kodama
Tatsuhiko
T
Siomi
Haruhiko
H
Siomi
Mikiko C
MC
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D030562:Databases, Protein; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D005786:Gene Expression Regulation; D013058:Mass Spectrometry / Q000379:methods; D026901:Membrane Transport Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D011496:Protein Methyltransferases / Q000378:metabolism*; D011484:Protein-Arginine N-Methyltransferases; D034622:RNA Interference; D017386:Sequence Homology, Amino Acid
D029721:Drosophila Proteins; D026901:Membrane Transport Proteins; C487735:dimethylarginine; C071597:tud protein, Drosophila; D001120:Arginine; D011496:Protein Methyltransferases; D011484:Protein-Arginine N-Methyltransferases; C512734:csul protein, Drosophila