Cargando…

A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila

BACKGROUND: Neuromuscular (NM) synaptogenesis is a tightly regulated process. We previously showed that in flies, Drosophila Nedd4 (dNedd4/dNedd4S) is required for proper NM synaptogenesis by promoting endocytosis of commissureless from the muscle surface, a pre-requisite step for muscle innervation...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhong, Yunan, Shtineman-Kotler, Alina, Nguyen, Leo, Iliadi, Konstantin G., Boulianne, Gabrielle L., Rotin, Daniela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3215714/
https://www.ncbi.nlm.nih.gov/pubmed/22110599
http://dx.doi.org/10.1371/journal.pone.0027007
_version_ 1782216425438445568
author Zhong, Yunan
Shtineman-Kotler, Alina
Nguyen, Leo
Iliadi, Konstantin G.
Boulianne, Gabrielle L.
Rotin, Daniela
author_facet Zhong, Yunan
Shtineman-Kotler, Alina
Nguyen, Leo
Iliadi, Konstantin G.
Boulianne, Gabrielle L.
Rotin, Daniela
author_sort Zhong, Yunan
collection PubMed
description BACKGROUND: Neuromuscular (NM) synaptogenesis is a tightly regulated process. We previously showed that in flies, Drosophila Nedd4 (dNedd4/dNedd4S) is required for proper NM synaptogenesis by promoting endocytosis of commissureless from the muscle surface, a pre-requisite step for muscle innervation. DNedd4 is an E3 ubiquitin ligase comprised of a C2-WW(x3)-Hect domain architecture, which includes several splice isoforms, the most prominent ones are dNedd4-short (dNedd4S) and dNedd4-long (dNedd4Lo). METHODOLOGY/PRINCIPAL FINDINGS: We show here that while dNedd4S is essential for NM synaptogenesis, the dNedd4Lo isoform inhibits this process and causes lethality. Our results reveal that unlike dNedd4S, dNedd4Lo cannot rescue the lethality of dNedd4 null (DNedd4(T121FS)) flies. Moreover, overexpression of UAS-dNedd4Lo specifically in wildtype muscles leads to NM synaptogenesis defects, impaired locomotion and larval lethality. These negative effects of dNedd4Lo are ameliorated by deletion of two regions (N-terminus and Middle region) unique to this isoform, and by inactivating the catalytic activity of dNedd4Lo, suggesting that these unique regions, as well as catalytic activity, are responsible for the inhibitory effects of dNedd4Lo on synaptogenesis. In accord with these findings, we demonstrate by sqRT-PCR an increase in dNedd4S expression relative to the expression of dNedd4Lo during embryonic stages when synaptogenesis takes place. CONCLUSION/SIGNIFICANCE: Our studies demonstrate that splice isoforms of the same dNedd4 gene can lead to opposite effects on NM synaptogenesis.
format Online
Article
Text
id pubmed-3215714
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-32157142011-11-21 A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila Zhong, Yunan Shtineman-Kotler, Alina Nguyen, Leo Iliadi, Konstantin G. Boulianne, Gabrielle L. Rotin, Daniela PLoS One Research Article BACKGROUND: Neuromuscular (NM) synaptogenesis is a tightly regulated process. We previously showed that in flies, Drosophila Nedd4 (dNedd4/dNedd4S) is required for proper NM synaptogenesis by promoting endocytosis of commissureless from the muscle surface, a pre-requisite step for muscle innervation. DNedd4 is an E3 ubiquitin ligase comprised of a C2-WW(x3)-Hect domain architecture, which includes several splice isoforms, the most prominent ones are dNedd4-short (dNedd4S) and dNedd4-long (dNedd4Lo). METHODOLOGY/PRINCIPAL FINDINGS: We show here that while dNedd4S is essential for NM synaptogenesis, the dNedd4Lo isoform inhibits this process and causes lethality. Our results reveal that unlike dNedd4S, dNedd4Lo cannot rescue the lethality of dNedd4 null (DNedd4(T121FS)) flies. Moreover, overexpression of UAS-dNedd4Lo specifically in wildtype muscles leads to NM synaptogenesis defects, impaired locomotion and larval lethality. These negative effects of dNedd4Lo are ameliorated by deletion of two regions (N-terminus and Middle region) unique to this isoform, and by inactivating the catalytic activity of dNedd4Lo, suggesting that these unique regions, as well as catalytic activity, are responsible for the inhibitory effects of dNedd4Lo on synaptogenesis. In accord with these findings, we demonstrate by sqRT-PCR an increase in dNedd4S expression relative to the expression of dNedd4Lo during embryonic stages when synaptogenesis takes place. CONCLUSION/SIGNIFICANCE: Our studies demonstrate that splice isoforms of the same dNedd4 gene can lead to opposite effects on NM synaptogenesis. Public Library of Science 2011-11-14 /pmc/articles/PMC3215714/ /pubmed/22110599 http://dx.doi.org/10.1371/journal.pone.0027007 Text en Zhong et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zhong, Yunan
Shtineman-Kotler, Alina
Nguyen, Leo
Iliadi, Konstantin G.
Boulianne, Gabrielle L.
Rotin, Daniela
A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title_full A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title_fullStr A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title_full_unstemmed A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title_short A Splice Isoform of DNedd4, DNedd4-Long, Negatively Regulates Neuromuscular Synaptogenesis and Viability in Drosophila
title_sort splice isoform of dnedd4, dnedd4-long, negatively regulates neuromuscular synaptogenesis and viability in drosophila
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3215714/
https://www.ncbi.nlm.nih.gov/pubmed/22110599
http://dx.doi.org/10.1371/journal.pone.0027007
work_keys_str_mv AT zhongyunan aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT shtinemankotleralina aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT nguyenleo aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT iliadikonstanting aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT bouliannegabriellel aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT rotindaniela aspliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT zhongyunan spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT shtinemankotleralina spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT nguyenleo spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT iliadikonstanting spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT bouliannegabriellel spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila
AT rotindaniela spliceisoformofdnedd4dnedd4longnegativelyregulatesneuromuscularsynaptogenesisandviabilityindrosophila