Cargando…

Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell

The spliceosome, responsible for all mature protein-coding transcripts of eukaryotic intron-containing genes, consists of small uridine-rich nuclear ribonucleoproteins (UsnRNPs). The assembly of UsnRNPs depends, on one hand, on the arginine methylation of Sm proteins catalyzed by the PRMT5 complex....

Descripción completa

Detalles Bibliográficos
Autores principales: Esser, Lea Marie, Schmitz, Katharina, Hillebrand, Frank, Erkelenz, Steffen, Schaal, Heiner, Stork, Björn, Grimmler, Matthias, Wesselborg, Sebastian, Peter, Christoph
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Research Network of Computational and Structural Biotechnology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034211/
https://www.ncbi.nlm.nih.gov/pubmed/36968021
http://dx.doi.org/10.1016/j.csbj.2023.03.015
_version_ 1784911162840711168
author Esser, Lea Marie
Schmitz, Katharina
Hillebrand, Frank
Erkelenz, Steffen
Schaal, Heiner
Stork, Björn
Grimmler, Matthias
Wesselborg, Sebastian
Peter, Christoph
author_facet Esser, Lea Marie
Schmitz, Katharina
Hillebrand, Frank
Erkelenz, Steffen
Schaal, Heiner
Stork, Björn
Grimmler, Matthias
Wesselborg, Sebastian
Peter, Christoph
author_sort Esser, Lea Marie
collection PubMed
description The spliceosome, responsible for all mature protein-coding transcripts of eukaryotic intron-containing genes, consists of small uridine-rich nuclear ribonucleoproteins (UsnRNPs). The assembly of UsnRNPs depends, on one hand, on the arginine methylation of Sm proteins catalyzed by the PRMT5 complex. On the other hand, it depends on the phosphorylation of the PRMT5 subunit pICln by the Uncoordinated Like Kinase 1 (ULK1). In consequence, phosphorylation of pICln affects the stability of the UsnRNP assembly intermediate, the so-called 6 S complex. The detailed mechanisms of phosphorylation-dependent integrity and subsequent UsnRNP assembly of the 6 S complex in vivo have not yet been analyzed. By using a phospho-specific antibody against ULK1-dependent phosphorylation sites of pICln, we visualize the intracellular distribution of phosphorylated pICln. Furthermore, we detect the colocaliphosphor-pICln1 with phospho-pICln by size-exclusion chromatography and immunofluorescence techniques. We also show that phosphorylated pICln is predominantly present in the 6 S complex. The addition of ULK1 to in vitro produced 6 S complex, as well as the reconstitution of ULK1 in ULK1-deficient cells, increases the efficiency of snRNP biogenesis. Accordingly, inhibition of ULK1 and the associated decreased pICln phosphorylation lead to accumulation of the 6 S complex and reduction in the spliceosomal activity of the cell.
format Online
Article
Text
id pubmed-10034211
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Research Network of Computational and Structural Biotechnology
record_format MEDLINE/PubMed
spelling pubmed-100342112023-03-24 Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell Esser, Lea Marie Schmitz, Katharina Hillebrand, Frank Erkelenz, Steffen Schaal, Heiner Stork, Björn Grimmler, Matthias Wesselborg, Sebastian Peter, Christoph Comput Struct Biotechnol J Research Article The spliceosome, responsible for all mature protein-coding transcripts of eukaryotic intron-containing genes, consists of small uridine-rich nuclear ribonucleoproteins (UsnRNPs). The assembly of UsnRNPs depends, on one hand, on the arginine methylation of Sm proteins catalyzed by the PRMT5 complex. On the other hand, it depends on the phosphorylation of the PRMT5 subunit pICln by the Uncoordinated Like Kinase 1 (ULK1). In consequence, phosphorylation of pICln affects the stability of the UsnRNP assembly intermediate, the so-called 6 S complex. The detailed mechanisms of phosphorylation-dependent integrity and subsequent UsnRNP assembly of the 6 S complex in vivo have not yet been analyzed. By using a phospho-specific antibody against ULK1-dependent phosphorylation sites of pICln, we visualize the intracellular distribution of phosphorylated pICln. Furthermore, we detect the colocaliphosphor-pICln1 with phospho-pICln by size-exclusion chromatography and immunofluorescence techniques. We also show that phosphorylated pICln is predominantly present in the 6 S complex. The addition of ULK1 to in vitro produced 6 S complex, as well as the reconstitution of ULK1 in ULK1-deficient cells, increases the efficiency of snRNP biogenesis. Accordingly, inhibition of ULK1 and the associated decreased pICln phosphorylation lead to accumulation of the 6 S complex and reduction in the spliceosomal activity of the cell. Research Network of Computational and Structural Biotechnology 2023-03-16 /pmc/articles/PMC10034211/ /pubmed/36968021 http://dx.doi.org/10.1016/j.csbj.2023.03.015 Text en © 2023 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Esser, Lea Marie
Schmitz, Katharina
Hillebrand, Frank
Erkelenz, Steffen
Schaal, Heiner
Stork, Björn
Grimmler, Matthias
Wesselborg, Sebastian
Peter, Christoph
Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title_full Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title_fullStr Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title_full_unstemmed Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title_short Phosphorylation of pICln by the autophagy activating kinase ULK1 regulates snRNP biogenesis and splice activity of the cell
title_sort phosphorylation of picln by the autophagy activating kinase ulk1 regulates snrnp biogenesis and splice activity of the cell
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10034211/
https://www.ncbi.nlm.nih.gov/pubmed/36968021
http://dx.doi.org/10.1016/j.csbj.2023.03.015
work_keys_str_mv AT esserleamarie phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT schmitzkatharina phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT hillebrandfrank phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT erkelenzsteffen phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT schaalheiner phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT storkbjorn phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT grimmlermatthias phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT wesselborgsebastian phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell
AT peterchristoph phosphorylationofpiclnbytheautophagyactivatingkinaseulk1regulatessnrnpbiogenesisandspliceactivityofthecell