Next generation organelles: Structure and role of germ granules in the germline

Gao, Ming; Arkov, Alexey L.

doi:10.1002/mrd.22115

Cited by 76 publications

(78 citation statements)

References 159 publications

(178 reference statements)

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“…Bombyx Vasa associates with the Piwi proteins Siwi and Ago3 (Xiol et al 2014;Nishida et al 2015), two major players in the Ping-Pong cycle in the germ plasm (Gao and Arkov 2013;Luteijn and Ketting 2013;Sato and Siomi 2013;Czech and Hannon 2016). Within the Ping-Pong cycle, Siwi is loaded with piRNAs, and the complex binds and cleaves transposon mRNAs in an orientation antisense to piRNAs.…”

Section: Discussionmentioning

confidence: 99%

“…During Drosophila oogenesis, Vasa localizes to two functionally distinct compartments in the egg chamber: the germ plasm in oocytes and the nuage in nurse cells (Mahowald 2001;Gao and Arkov 2013). Nurse cells are transcriptionally active and provide the growing oocyte with RNAs and proteins required for oocyte development and patterning of the future embryo (Johnstone and Lasko 2001).…”

mentioning

confidence: 99%

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The LOTUS domain is a conserved DEAD-box RNA helicase regulator essential for the recruitment of Vasa to the germ plasm and nuage

2017

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DEAD-box RNA helicases play important roles in a wide range of metabolic processes. Regulatory proteins can stimulate or block the activity of DEAD-box helicases. Here, we show that LOTUS (Limkain, Oskar, and Tudor containing proteins 5 and 7) domains present in the germline proteins Oskar, TDRD5 (Tudor domain-containing 5), and TDRD7 bind and stimulate the germline-specific DEAD-box RNA helicase Vasa. Our crystal structure of the LOTUS domain of Oskar in complex with the C-terminal RecA-like domain of Vasa reveals that the LOTUS domain occupies a surface on a DEAD-box helicase not implicated previously in the regulation of the enzyme's activity. We show that, in vivo, the localization of Drosophila Vasa to the nuage and germ plasm depends on its interaction with LOTUS domain proteins. The binding and stimulation of Vasa DEAD-box helicases by LOTUS domains are widely conserved.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

The LOTUS domain is a conserved DEAD-box RNA helicase regulator essential for the recruitment of Vasa to the germ plasm and nuage

2017

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“…PRMT5 catalyzes the synthesis of symmetrical dimethylated arginines (sDMA) on SmB, Vasa and PIWI homologs (Blackwell and Ceman, 2012). These sDMA modifications are binding sites for Tudor domain-containing proteins and contribute to the aggregation and function of these granules (Gao and Arkov, 2012;Siomi et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

PIWI homologs mediate Histone H4 mRNA localization to planarian chromatoid bodies

et al. 2014

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The well-known regenerative abilities of planarian flatworms are attributed to a population of adult stem cells called neoblasts that proliferate and differentiate to produce all cell types. A characteristic feature of neoblasts is the presence of large cytoplasmic ribonucleoprotein granules named chromatoid bodies, the function of which has remained largely elusive. This study shows that histone mRNAs are a common component of chromatoid bodies. Our experiments also demonstrate that accumulation of histone mRNAs, which is typically restricted to the S phase of eukaryotic cells, is extended during the cell cycle of neoblasts. The planarian PIWI homologs SMEDWI-1 and SMEDWI-3 are required for proper localization of germinal histone H4 (gH4) mRNA to chromatoid bodies. The association between histone mRNA and chromatoid body components extends beyond gH4 mRNA, since transcripts of other core histone genes were also found in these structures. Additionally, piRNAs corresponding to loci of every core histone type have been identified. Altogether, this work provides evidence that links PIWI proteins and chromatoid bodies to histone mRNA regulation in planarian stem cells. The molecular similarities between neoblasts and undifferentiated cells of other organisms raise the possibility that PIWI proteins might also regulate histone mRNAs in stem cells and germ cells of other metazoans.

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“…Tud is a germline protein which is essential for germline development [4][5][6][7] . This protein contains 11 Tud domains known to interact with methylated arginines or lysines of other polypeptides [8][9][10] . Previously, we have generated a transgenic Drosophila line which expresses HA-tagged functional Tud 5 and therefore, specific anti-HA antibody is used to pull down Tud complex after crosslinking.…”

Section: Introductionmentioning

confidence: 99%

An <em>in vivo</em> Crosslinking Approach to Isolate Protein Complexes From <em>Drosophila</em> Embryos

Gao

McCluskey

Loganathan

et al. 2014

JoVE

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Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to assemble. Therefore, to identify these functional protein complexes, it is important to stabilize them in vivo before cell lysis and subsequent purification. Here we describe a method used to isolate large bona fide protein complexes from Drosophila embryos. This method is based on embryo permeabilization and stabilization of the complexes inside the embryos by in vivo crosslinking using a low concentration of formaldehyde, which can easily cross the cell membrane. Subsequently, the protein complex of interest is immunopurified followed by gel purification and analyzed by mass spectrometry. We illustrate this method using purification of a Tudor protein complex, which is essential for germline development. Tudor is a large protein, which contains multiple Tudor domains -small modules that interact with methylated arginines or lysines of target proteins. This method can be adapted for isolation of native protein complexes from different organisms and tissues.

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Next generation organelles: Structure and role of germ granules in the germline

Cited by 76 publications

References 159 publications

The LOTUS domain is a conserved DEAD-box RNA helicase regulator essential for the recruitment of Vasa to the germ plasm and nuage

The LOTUS domain is a conserved DEAD-box RNA helicase regulator essential for the recruitment of Vasa to the germ plasm and nuage

PIWI homologs mediate Histone H4 mRNA localization to planarian chromatoid bodies

An <em>in vivo</em> Crosslinking Approach to Isolate Protein Complexes From <em>Drosophila</em> Embryos

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