Half Pint/Puf68 is required for negative regulation of splicing by the SR factor Transformer2

Wang, Shanzhi; Wagner, Eric J.; Mattox, William

doi:10.4161/rna.25645

Cited by 7 publications

(6 citation statements)

References 49 publications

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“…To investigate whether Hfp plays a direct role on Kdm3 splicing, RNA immunoprecipitation (RIP) experiments were carried out using Hfp-GFP expressing flies and anti-GFP antibodies followed by RT-qPCR analyses. As expected, RNA from known targets of hfp such as tra2 22 and otu 18 were significantly enriched when compared to a control gene (eEF5) (Fig. 2f).…”

Section: Figuresupporting

confidence: 79%

The histone demethylase KDM3 prevents auto-immune piRNAs production in Drosophila

Casier

Autaa

Guegen

et al. 2022

Preprint

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In animals, genome integrity of the germ line is protected from transposable element (TE) activity by small, non-coding, dedicated RNAs acting as an immune system against TEs, and called PIWI-interacting RNAs (piRNAs). In Drosophila, the production of piRNAs is initiated from heterochromatic loci containing remnants of TEs and enriched in histone H3 trimethylated on lysine 9 (H3K9me3). These loci, called piRNA clusters, constitute a memory of past TE invasions. Little is known about how piRNA clusters are genetically defined. Using a genetic screen combined with a bimodal epigenetic state piRNA cluster (BX2), we identified the splicing factor Half pint (Hfp) and the histone demethylase KDM3 as being able to prevent BX2 piRNA production. Furthermore, we showed that Hfp is needed to splice Kdm3 transcripts. Germline expression of Kdm3 coding sequence (splicing-independent) rescued the hfp germline knock-down (GLKD) effect demonstrating that Kdm3 is sufficient to prevent BX2 piRNA production. Our data revealed that in the absence of Kdm3, dozens of gene-containing regions become bona fide germinal dual strand piRNA clusters. Indeed, they produce piRNAs originating from both DNA strands, become transcribed in a Moonshiner-dependent manner and enriched in di-and tri-methylation of lysine 9 of histone H3 (H3K9me2/3) and in Rhino, an HP1-like protein. Eggs laid by Kdm3 GLKD females do not hatch and show developmental defects phenocopying loss of function of genes included into the new piRNA clusters, suggesting an inheritance of functional ovarian "auto-immune" piRNAs. Our results demonstrate that some gene-containing regions are actively prevented for piRNA production by proteins that counteract piRNA cluster emergence. Hence, a non-piRNA-producing state is therefore not a "by default" state but rather a cellular lock that is actively controlled for some genomic loci.

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Section: Figuresupporting

confidence: 79%

The histone demethylase KDM3 prevents auto-immune piRNAs production in Drosophila

Casier

Autaa

Guegen

et al. 2022

Preprint

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“…In D. melanogaster Tra-2 (a SR protein), together with Tra act as a splicing activator for splicing of dsx pre-mRNA into female mode (Nagoshi et al, 1988). Tra-2, together with Puf68, act as splicing repressor of male-specific exon of its own pre-mRNA in testis (Wang et al, 2013). …”

Section: Discussionmentioning

confidence: 99%

Tribolium castaneum Transformer-2 regulates sex determination and development in both males and females

Shukla

Palli

2013

Insect Biochemistry and Molecular Biology

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Tribolium castaneum Transformer (TcTra) is essential for female sex determination and maintenance through the regulation of sex-specific splicing of doublesex (dsx) pre-mRNA. In females, TcTra also regulates the sex-specific splicing of its own pre-mRNA to ensure continuous production of functional Tra protein. Transformer protein is absent in males and hence dsx pre-mRNA is spliced in a default mode. The mechanisms by which males inhibit the production of functional Tra protein are not known. Here, we report on functional characterization of transformer-2 (tra-2) gene (an ortholog of Drosophila transformer-2) in T. castaneum. RNA interference-mediated knockdown in the expression of gene coding for tra-2 in female pupae or adults resulted in the production of male-specific isoform of dsx and both female and male isoforms of tra suggesting that Tra-2 is essential for the female-specific splicing of tra and dsx pre-mRNAs. Interestingly, knockdown of tra-2 in males did not affect the splicing of dsx but resulted in the production of both female and male isoforms of tra suggesting that Tra-2 suppresses female-specific splicing of tra pre-mRNA in males. This dual regulation of sex-specific splicing of tra pre-mRNA ensures a tight regulation of sex determination and maintenance. These data suggest a critical role for Tra-2 in suppression of female sex determination cascade in males. In addition, RNAi studies showed that Tra-2 is also required for successful embryonic and larval development in both sexes.

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“…This gene is differentially expressed between adult virgin males and females (McIntyre et al , 2006), and a loss-of-function mutation leads to malformed sex combs (Quinn et al , 2004). The product of pUf68 is known to regulate alternative splicing (Van Buskirk and Schupbach, 2002), and is required for splicing of the M1 intron from the tra2 primary transcript, although it does not appear to contribute to Tra2 control of dsx splicing (Wang et al , 2013). scrib is present within QTL Q3 on chromosome 3 and, similar to disco-r , has been shown to have lower gene expression in male forelegs compared with female forelegs (Barmina et al , 2005).…”

Section: Discussionmentioning

confidence: 99%

Genetic dissection of intraspecific variation in a male-specific sexual trait in Drosophila melanogaster

2016

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An open question in evolutionary biology is the relationship between standing variation for a trait and the variation that leads to interspecific divergence. By identifying loci underlying phenotypic variation in intra- and interspecific crosses we can determine the extent to which polymorphism and divergence are controlled by the same genomic regions. Sexual traits provide abundant examples of morphological and behavioral diversity within and among species, and here we leverage variation in the Drosophila sex comb to address this question. The sex comb is an array of modified bristles or ‘teeth' present on the male forelegs of several Drosophilid species. Males use the comb to grasp females during copulation, and ablation experiments have shown that males lacking comb teeth typically fail to mate. We measured tooth number in >700 genotypes derived from a multiparental advanced-intercross population, mapping three moderate-effect loci contributing to trait heritability. Two quantitative trait loci (QTLs) coincide with previously identified intra- and interspecific sex comb QTL, but such overlap can be explained by chance alone, in part because of the broad swathes of the genome implicated by earlier, low-resolution QTL scans. Our mapped QTL regions encompass 70–124 genes, but do not include those genes known to be involved in developmental specification of the comb. Nonetheless, we identified plausible candidates within all QTL intervals, and used RNA interference to validate effects at four loci. Notably, TweedleS expression knockdown substantially reduces tooth number. The genes we highlight are strong candidates to harbor segregating, functional variants contributing to sex comb tooth number.

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Half Pint/Puf68 is required for negative regulation of splicing by the SR factor Transformer2

Cited by 7 publications

References 49 publications

The histone demethylase KDM3 prevents auto-immune piRNAs production in Drosophila

The histone demethylase KDM3 prevents auto-immune piRNAs production in Drosophila

Tribolium castaneum Transformer-2 regulates sex determination and development in both males and females

Genetic dissection of intraspecific variation in a male-specific sexual trait in Drosophila melanogaster

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