Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes
Spliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of t...
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Taylor & Francis Group
2024-12-01
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| Series: | RNA Biology |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/15476286.2024.2348896 |
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| author | Kateřina Abrhámová Martina Groušlová Anna Valentová Xinxin Hao Beidong Liu Martin Převorovský Ondřej Gahura František Půta Per Sunnerhagen Petr Folk |
| author_facet | Kateřina Abrhámová Martina Groušlová Anna Valentová Xinxin Hao Beidong Liu Martin Převorovský Ondřej Gahura František Půta Per Sunnerhagen Petr Folk |
| author_sort | Kateřina Abrhámová |
| collection | DOAJ |
| description | Spliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of the genetic interaction network of the truncated allele of PRP45 (prp45(1–169)) using synthetic genetic array technology and found chromatin remodellers and modifiers as an enriched category. In agreement with related studies, H2A.Z-encoding HTZ1, and the components of SWR1, INO80, and SAGA complexes represented prominent interactors, with htz1 conferring the strongest growth defect. Because the truncation of Prp45 disproportionately affected low copy number transcripts of intron-containing genes, we prepared strains carrying intronless versions of SRB2, VPS75, or HRB1, the most affected cases with transcription-related function. Intron removal from SRB2, but not from the other genes, partly repaired some but not all the growth phenotypes identified in the genetic screen. The interaction of prp45(1–169) and htz1Δ was detectable even in cells with SRB2 intron deleted (srb2Δi). The less truncated variant, prp45(1–330), had a synthetic growth defect with htz1Δ at 16°C, which also persisted in the srb2Δi background. Moreover, htz1Δ enhanced prp45(1–330) dependent pre-mRNA hyper-accumulation of both high and low efficiency splicers, genes ECM33 and COF1, respectively. We conclude that while the expression defects of low expression intron-containing genes contribute to the genetic interactome of prp45(1–169), the genetic interactions between prp45 and htz1 alleles demonstrate the sensitivity of spliceosome assembly, delayed in prp45(1–169), to the chromatin environment. |
| format | Article |
| id | doaj-art-2f96b6e1b3c9407b985d3b7740507fc0 |
| institution | DOAJ |
| issn | 1547-6286 1555-8584 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
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| series | RNA Biology |
| spelling | doaj-art-2f96b6e1b3c9407b985d3b7740507fc02025-08-20T02:50:08ZengTaylor & Francis GroupRNA Biology1547-62861555-85842024-12-0121154355910.1080/15476286.2024.2348896Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypesKateřina Abrhámová0Martina Groušlová1Anna Valentová2Xinxin Hao3Beidong Liu4Martin Převorovský5Ondřej Gahura6František Půta7Per Sunnerhagen8Petr Folk9Department of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicDepartment of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicDepartment of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicDepartment of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, SwedenDepartment of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicInstitute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice, Czech RepublicDepartment of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicDepartment of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, SwedenDepartment of Cell Biology, Faculty of Science, Charles University, Praha, Czech RepublicSpliceosome assembly contributes an important but incompletely understood aspect of splicing regulation. Prp45 is a yeast splicing factor which runs as an extended fold through the spliceosome, and which may be important for bringing its components together. We performed a whole genome analysis of the genetic interaction network of the truncated allele of PRP45 (prp45(1–169)) using synthetic genetic array technology and found chromatin remodellers and modifiers as an enriched category. In agreement with related studies, H2A.Z-encoding HTZ1, and the components of SWR1, INO80, and SAGA complexes represented prominent interactors, with htz1 conferring the strongest growth defect. Because the truncation of Prp45 disproportionately affected low copy number transcripts of intron-containing genes, we prepared strains carrying intronless versions of SRB2, VPS75, or HRB1, the most affected cases with transcription-related function. Intron removal from SRB2, but not from the other genes, partly repaired some but not all the growth phenotypes identified in the genetic screen. The interaction of prp45(1–169) and htz1Δ was detectable even in cells with SRB2 intron deleted (srb2Δi). The less truncated variant, prp45(1–330), had a synthetic growth defect with htz1Δ at 16°C, which also persisted in the srb2Δi background. Moreover, htz1Δ enhanced prp45(1–330) dependent pre-mRNA hyper-accumulation of both high and low efficiency splicers, genes ECM33 and COF1, respectively. We conclude that while the expression defects of low expression intron-containing genes contribute to the genetic interactome of prp45(1–169), the genetic interactions between prp45 and htz1 alleles demonstrate the sensitivity of spliceosome assembly, delayed in prp45(1–169), to the chromatin environment.https://www.tandfonline.com/doi/10.1080/15476286.2024.2348896Synthetic genetic array analysisspliceosome assemblyco-transcriptional splicingchromatin modifiersH2A.Z |
| spellingShingle | Kateřina Abrhámová Martina Groušlová Anna Valentová Xinxin Hao Beidong Liu Martin Převorovský Ondřej Gahura František Půta Per Sunnerhagen Petr Folk Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes RNA Biology Synthetic genetic array analysis spliceosome assembly co-transcriptional splicing chromatin modifiers H2A.Z |
| title | Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes |
| title_full | Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes |
| title_fullStr | Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes |
| title_full_unstemmed | Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes |
| title_short | Truncating the spliceosomal ‘rope protein’ Prp45 results in Htz1 dependent phenotypes |
| title_sort | truncating the spliceosomal rope protein prp45 results in htz1 dependent phenotypes |
| topic | Synthetic genetic array analysis spliceosome assembly co-transcriptional splicing chromatin modifiers H2A.Z |
| url | https://www.tandfonline.com/doi/10.1080/15476286.2024.2348896 |
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