Blom7α is a novel heterogeneous nuclear ribonucleoprotein K homology domain protein involved in pre-mRNA splicing that interacts with SNEVPrp19-Pso4

Johannes Grillari, Marlies Löscher, Marco Denegri, Kiseok Lee, Klaus Fortschegger, Frank Eisenhaber, Paul Ajuh, Angus I. Lamond, Hermann Katinger, Regina Grillari-Voglauer

Research output: Contribution to journalArticle

Abstract

The removal of introns from pre-mRNA is performed by the spliceosome that stepwise assembles on the pre-mRNA before performing two catalytic steps. The spliceosome-associated CDC5L-SNEVPrp19-Pso4 complex is implicated in activation of the second catalytic step of pre-mRNA splicing, and one of its members, SNEVPrp19-Pso4, is also implicated in spliceosome assembly. To identify interaction partners of SNEVPrp19-Pso4, we have performed yeast two-hybrid screenings. Among the putative binding partners was a so far uncharacterized protein carrying two heterogeneous nuclear ribonucleoprotein K homology domains that we termed Blom7α. Blom7α is expressed in all tissues tested, and at least three splice variants exist. After confirming direct and physical interaction of SNEV and Blom7α, we investigated if it plays a functional role during pre-mRNA splicing. Indeed, Blom7α co-localizes and co-precipitates with splicing factors and pre-mRNA and is present in affinity-purified spliceosomes. More importantly, addition of Blom7αto HeLa nuclear extracts increased splicing activity in a dose-dependent manner. Furthermore, we tested if Blom7α influences splice site selection using two different minigene constructs. Indeed, both 5′- as well as 3′-site selection was altered upon Blom7α overexpression. Thus we suggest that Blom7α is a novel splicing factor of the K homology domain family that might be implicated in alternative splicing by helping to position the CDC5L-SNEVPrp19-Pso4 complex at the splice sites.

Original languageEnglish
Pages (from-to)29193-29204
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number42
DOIs
Publication statusPublished - Oct 16 2009

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Heterogeneous-Nuclear Ribonucleoprotein K
RNA Precursors
Spliceosomes
Site selection
Proteins
Alternative Splicing
Yeast
Introns
Precipitates
Screening
Yeasts
Chemical activation
Protein Domains
Tissue

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

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Blom7α is a novel heterogeneous nuclear ribonucleoprotein K homology domain protein involved in pre-mRNA splicing that interacts with SNEVPrp19-Pso4 . / Grillari, Johannes; Löscher, Marlies; Denegri, Marco; Lee, Kiseok; Fortschegger, Klaus; Eisenhaber, Frank; Ajuh, Paul; Lamond, Angus I.; Katinger, Hermann; Grillari-Voglauer, Regina.

In: Journal of Biological Chemistry, Vol. 284, No. 42, 16.10.2009, p. 29193-29204.

Research output: Contribution to journalArticle

Grillari, J, Löscher, M, Denegri, M, Lee, K, Fortschegger, K, Eisenhaber, F, Ajuh, P, Lamond, AI, Katinger, H & Grillari-Voglauer, R 2009, 'Blom7α is a novel heterogeneous nuclear ribonucleoprotein K homology domain protein involved in pre-mRNA splicing that interacts with SNEVPrp19-Pso4 ', Journal of Biological Chemistry, vol. 284, no. 42, pp. 29193-29204. https://doi.org/10.1074/jbc.M109.036632
Grillari, Johannes ; Löscher, Marlies ; Denegri, Marco ; Lee, Kiseok ; Fortschegger, Klaus ; Eisenhaber, Frank ; Ajuh, Paul ; Lamond, Angus I. ; Katinger, Hermann ; Grillari-Voglauer, Regina. / Blom7α is a novel heterogeneous nuclear ribonucleoprotein K homology domain protein involved in pre-mRNA splicing that interacts with SNEVPrp19-Pso4 . In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 42. pp. 29193-29204.
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AU - Denegri, Marco

AU - Lee, Kiseok

AU - Fortschegger, Klaus

AU - Eisenhaber, Frank

AU - Ajuh, Paul

AU - Lamond, Angus I.

AU - Katinger, Hermann

AU - Grillari-Voglauer, Regina

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