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Identification of MVH target mRNA

ABSTRACT: During mammalian spermatogenesis the mouse VASA homolog (MVH), a germ cell-specific DEAD-box type RNA binding protein, localizes in a germline-specific RNA granule termed the chromatoid body (CB). Genetic analyses have revealed that MVH is essential to progress through spermatogenesis, although the molecular mechanisms of its function remain elusive. We found that the acetyltransferase HAT1 and its cofactor, p46, are specifically co-localized with MVH in the CB and acetylate MVH at Lys-405, leading to inactivation of its RNA binding activity. Notably, the acetylation is developmentally regulated, paralleling the temporally regulated co-localization of HAT1 and p46 in the CB. We have identified 858 mRNAs as MVH targets, a large proportion of which correspond to previously identified translationally arrested genes. Importantly, eIF4B mRNA, a target of MVH, is selectively released from MVH-RNP when MVH is acetylated, paralleling an increase in eIF4B protein. These findings reveal a novel signaling pathway that links acetylation to RNA processing in the control of spermatogenesis. IMVH target mRNA was purified from MVH immuno precipitated complex, followed by RNA purification and identified by DNA chip. In this study, we identified more than 800 MVH target mRNA in testis.

ORGANISM(S): Mus musculus

SUBMITTER: Ippei Nagamori

PROVIDER: E-GEOD-30144 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Identification of MVH target mRNA

Project description:During mammalian spermatogenesis the mouse VASA homolog (MVH), a germ cell-specific DEAD-box type RNA binding protein, localizes in a germline-specific RNA granule termed the chromatoid body (CB). Genetic analyses have revealed that MVH is essential to progress through spermatogenesis, although the molecular mechanisms of its function remain elusive. We found that the acetyltransferase HAT1 and its cofactor, p46, are specifically co-localized with MVH in the CB and acetylate MVH at Lys-405, leading to inactivation of its RNA binding activity. Notably, the acetylation is developmentally regulated, paralleling the temporally regulated co-localization of HAT1 and p46 in the CB. We have identified 858 mRNAs as MVH targets, a large proportion of which correspond to previously identified translationally arrested genes. Importantly, eIF4B mRNA, a target of MVH, is selectively released from MVH-RNP when MVH is acetylated, paralleling an increase in eIF4B protein. These findings reveal a novel signaling pathway that links acetylation to RNA processing in the control of spermatogenesis.

2011-08-31 | GSE30144 | GEO

The Atlas of Chromatoid Body Components [RNA-seq]

Project description:The genome of male germ cells is actively transcribed during spermatogenesis to produce phase-specific protein coding mRNAs and a considerable amount of different non-coding RNAs. Ribonucleoprotein (RNP) granule-mediated RNA regulation provides a powerful means to secure the quality and correct expression of the requisite transcripts. Haploid spermatids are characterized by a unique, unusually large cytoplasmic granule, the chromatoid body (CB), that emerges during the switch between the meiotic and post-meiotic phases of spermatogenesis. To better understand the role of the CB in male germ cell differentiation, we isolated CBs from mouse testes and revealed its full RNA and protein composition. We showed that the CB is mainly composed of RNA-binding proteins and other proteins involved RNA regulation. The CB was loaded with RNA, including pachytene piRNAs, a diverse set of mRNAs and a number of uncharacterized long non-coding transcripts. The CB was demonstrated to accumulate nascent RNA during all the steps of round spermatid differentiation. Our results revealed the CB as a large germ cell-specific RNP platform that is involved in the control of the highly complex transcriptome of haploid male germ cells. Transcriptome profling of purified chromatoid body, each steps of chromatoid body purification, and sortred round spermatid cells

2014-01-24 | E-GEOD-54286 | biostudies-arrayexpress

The Atlas of Chromatoid Body Components [small RNA-seq]

Project description:The genome of male germ cells is actively transcribed during spermatogenesis to produce phase-specific protein coding mRNAs and a considerable amount of different non-coding RNAs. Ribonucleoprotein (RNP) granule-mediated RNA regulation provides a powerful means to secure the quality and correct expression of the requisite transcripts. Haploid spermatids are characterized by a unique, unusually large cytoplasmic granule, the chromatoid body (CB), that emerges during the switch between the meiotic and post-meiotic phases of spermatogenesis. To better understand the role of the CB in male germ cell differentiation, we isolated CBs from mouse testes and revealed its full RNA and protein composition. We showed that the CB is mainly composed of RNA-binding proteins and other proteins involved RNA regulation. The CB was loaded with RNA, including pachytene piRNAs, a diverse set of mRNAs and a number of uncharacterized long non-coding transcripts. The CB was demonstrated to accumulate nascent RNA during all the steps of round spermatid differentiation. Our results revealed the CB as a large germ cell -specific RNP platform that is involved in the control of the highly complex transcriptome of haploid male germ cells. Small RNA profiling of purified chromatoid body, each steps of chromatoid body purification, and sortred round spermatid cells

2014-01-24 | E-GEOD-54287 | biostudies-arrayexpress

Molecular role of RNA helicases MVH and TDRD9 in PIWI slicing-triggered mammalian piRNA biogenesis and function

Project description:PIWI-interacting RNAs (piRNAs) function in the nucleus and cytoplasm of animal germ cells to suppress mobile genetic elements. In the mouse male germline, biogenesis of MIWI2-bound nuclear piRNAs depends on endonuclease activity of cytosolic MILI, but the process is poorly understood. Here we use a mouse model expressing an artificial piRNA precursor to show that MILI slicing of the precursor generates a 16-nt by-product and a pre-piRNA intermediate that requires 3ʹ end processing to mature as a new piRNA. The ability to use the slicer products requires ATPase activity of the RNA helicase MVH, as the catalytic-dead Mvh mutant mice (Mvh-/KI) fail to convert the intermediate into piRNAs, abrogating biogenesis of MIWI2 piRNAs. This results in an early arrest in spermatogenesis and de-repression of transposons. Furthermore, the mutant MVH protein is dominant-negative (Mvh+/KI) as it causes a late-spermatogenic arrest by trapping complexes containing the mysterious pachytene piRNAs and slicer products, uncovering a role for the protein beyond the embryonic germline. In contrast, we find that the ATPase activity of TDRD9 is dispensable for piRNA biogenesis, but is essential for silencing by MIWI2. Our studies implicate distinct RNA helicases in specific steps along the mammalian nuclear piRNA pathway.

2017-06-20 | GSE95580 | GEO

The Atlas of Chromatoid Body Components [small RNA-seq]

Project description:The genome of male germ cells is actively transcribed during spermatogenesis to produce phase-specific protein coding mRNAs and a considerable amount of different non-coding RNAs. Ribonucleoprotein (RNP) granule-mediated RNA regulation provides a powerful means to secure the quality and correct expression of the requisite transcripts. Haploid spermatids are characterized by a unique, unusually large cytoplasmic granule, the chromatoid body (CB), that emerges during the switch between the meiotic and post-meiotic phases of spermatogenesis. To better understand the role of the CB in male germ cell differentiation, we isolated CBs from mouse testes and revealed its full RNA and protein composition. We showed that the CB is mainly composed of RNA-binding proteins and other proteins involved RNA regulation. The CB was loaded with RNA, including pachytene piRNAs, a diverse set of mRNAs and a number of uncharacterized long non-coding transcripts. The CB was demonstrated to accumulate nascent RNA during all the steps of round spermatid differentiation. Our results revealed the CB as a large germ cell -specific RNP platform that is involved in the control of the highly complex transcriptome of haploid male germ cells.

2014-01-24 | GSE54287 | GEO

The Atlas of Chromatoid Body Components [RNA-seq]

2014-01-24 | GSE54286 | GEO

Hat mutants

Project description:Transcriptomes from the hat1, hat2 and hat1 hat2 mutants in S288C yeast background.

2010-06-09 | E-GEOD-2434 | biostudies-arrayexpress

Hat mutants

Project description:Transcriptomes from the hat1, hat2 and hat1 hat2 mutants in S288C yeast background. Keywords: parallel sample

2005-07-23 | GSE2434 | GEO

HAT1 coordinates histone production and acetylation via H4 promoter binding

Project description:The energetic costs of duplicating chromatin are large and therefore likely depend on nutrient sensing checkpoints and metabolic inputs. By studying chromatin modifiers regulated by epithelial growth factor, we identified histone acetyltransferase 1 (HAT1) as an induced gene that enhances proliferation through coordinating histone production, acetylation and glucose metabolism. In addition to its canonical role as a cytoplasmic histone H4 acetyltransferase, we isolated a HAT1-containing complex bound specifically at promoters of H4 genes. HAT1-dependent transcription of H4 genes required an acetate-sensitive promoter element. HAT1 expression was critical for S-phase progression and maintenance of H3 lysine 9 acetylation at proliferation-associated genes, including histone genes. Therefore, these data describe a feed-forward circuit whereby HAT1 captures acetyl-groups on nascent histones and drives H4 production by chromatin binding to support chromatin replication and acetylation. These findings have important implications for human disease, since high HAT1 levels associate with poor outcomes across multiple cancer types.

2019-07-02 | GSE117472 | GEO

Hat1 WT and Hat1 KO MEFs analyzed to identify sites of acetylation that are dependent on Hat1

Project description:Hat1 WT and Hat1 KO MEFs were grown on both galactose and glucose. Abundance of acetylated peptides was compared between the samples.

2020-02-28 | PXD017362 | Pride

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