<i>Drosophila</i>Nedd4-long reduces Amphiphysin levels in muscles and leads to impaired T-tubule formation

Safi, Frozan; Shteiman-Kotler, Alina; Zhong, Yunan; Iliadi, Konstantin G.; Boulianne, Gabrielle L.; Rotin, Daniela

doi:10.1091/mbc.e15-06-0420

Cited by 5 publications

(13 citation statements)

References 37 publications

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“…It thus appears that the SH3 domain of SH3PX1 provides the sole binding interface to dNedd4Lo, a less common mode of interaction between a Nedd4 family member and its substrates. A similar phenomenon was observed with the dNedd4Lo⅐dAmph interaction (18). Interestingly, SNX9 was identified as a substrate of the Nedd4 family member Itch in melanoma cells, where the SNX9 SH3 domain binds a proline-rich region in Itch (29).…”

Section: Discussionsupporting

confidence: 74%

“…To identify interacting partners to the unique regions of dNedd4Lo, Drosophila embryo lysates were incubated with purified, GST-immobilized N-terminal and Mid regions and analyzed by MS. Two of the high-confidence interacting partners were identified as binding partners to dNedd4Lo were Drosophila amphiphysin (dAmph) and SH3PX1. dAmph, which binds to the unique N-terminal region of dNedd4Lo, was shown to be degraded by dNedd4Lo in the muscle and to regulate T-tubule formation and larval locomotion (18). Our screen also identified SH3PX1 as a binding partner to the dNedd4Lo unique Mid region.…”

Section: Neuromuscular Synaptogenesis (Nms)mentioning

confidence: 70%

“…Because previous studies implicated dNedd4Lo as a negative regulator of NMS and larval locomotion, we sought to determine how it regulates these processes through its interacting partners. dNedd4Lo was shown to genetically interact with dAmph in the body wall muscles, where it regulates T-tubule architecture and larval locomotion at least partially through dAmph degradation (18). In the current study we tested whether the dNedd4Lo⅐SH3PX1 interaction could help elucidate how dNedd4Lo regulates NMS and muscle innervation, which we previously showed to be impaired in dNedd4Lo (9).…”

Section: Discussionmentioning

confidence: 92%

“…We showed previously that dNedd4 is expressed throughout Drosophila muscles where it plays a role in T-tubule formation and is required for proper innervation of the SNb branch from muscle 13 to 12 (8,9,18). If dNedd4Lo regulates either of these processes via SH3PX1, we would expect SH3PX1 mutant larvae to display similar defects.…”

Section: Dnedd4lo Does Not Regulate Muscle T-tubule Architecture or Smentioning

confidence: 89%

“…We previously identified an interaction between dNedd4Lo-Mid and SH3PX1 using mass spectrometry (MS) ( Fig. 1A) (18). To validate the MS results, we first tested whether SH3PX1 co-immunoprecipitates (co-IP) with Nedd4Lo in cells.…”

Section: Sh3px1 Interacts With Dnedd4lomentioning

confidence: 95%

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Regulation of SH3PX1 by dNedd4-long at the Drosophila neuromuscular junction

Wasserman

Shteiman-Kotler

Harris

et al. 2019

Journal of Biological Chemistry

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Drosophila Nedd4 (dNedd4) is a HECT E3 ubiquitin ligase present in two major isoforms: short (dNedd4S) and long (dNedd4Lo), with the latter containing two unique regions (N terminus and Middle). Although dNedd4S promotes neuromuscular synaptogenesis (NMS), dNedd4Lo inhibits it and impairs larval locomotion. To explain how dNedd4Lo inhibits NMS, MS analysis was performed to find its binding partners and identified SH3PX1, which binds dNedd4Lo unique Middle region. SH3PX1 contains SH3, PX, and BAR domains and is present at neuromuscular junctions, where it regulates active zone ultrastructure and presynaptic neurotransmitter release. Here, we demonstrate direct binding of SH3PX1 to the dNedd4Lo Middle region (which contains a Pro-rich sequence) in vitro and in cells, via the SH3PX1-SH3 domain. In Drosophila S2 cells, dNedd4Lo overexpression reduces SH3PX1 levels at the cell periphery. In vivo overexpression of dNedd4Lo postsynaptically, but not pre-synaptically, reduces SH3PX1 levels at the subsynaptic reticulum and impairs neurotransmitter release. Unexpectedly, larvae that overexpress dNedd4Lo postsynaptically and are heterozygous for a null mutation in SH3PX1 display increased neurotransmission compared with dNedd4Lo or SH3PX1 mutant larvae alone, suggesting a compensatory effect from the remaining SH3PX1 allele. These results suggest a postsynaptic-specific regulation of SH3PX1 by dNedd4Lo.

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Section: Discussionsupporting

confidence: 74%

Section: Neuromuscular Synaptogenesis (Nms)mentioning

confidence: 70%

Section: Discussionmentioning

confidence: 92%

Section: Dnedd4lo Does Not Regulate Muscle T-tubule Architecture or Smentioning

confidence: 89%

Section: Sh3px1 Interacts With Dnedd4lomentioning

confidence: 95%

See 3 more Smart Citations

Regulation of SH3PX1 by dNedd4-long at the Drosophila neuromuscular junction

Wasserman

Shteiman-Kotler

Harris

et al. 2019

Journal of Biological Chemistry

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Lysine acetylation regulates the interaction between proteins and membranes

et al. 2021

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Lysine acetylation regulates the function of soluble proteins in vivo, yet it remains largely unexplored whether lysine acetylation regulates membrane protein function. Here, we use bioinformatics, biophysical analysis of recombinant proteins, live-cell fluorescent imaging and genetic manipulation of Drosophila to explore lysine acetylation in peripheral membrane proteins. Analysis of 50 peripheral membrane proteins harboring BAR, PX, C2, or EHD membrane-binding domains reveals that lysine acetylation predominates in membrane-interaction regions. Acetylation and acetylation-mimicking mutations in three test proteins, amphiphysin, EHD2, and synaptotagmin1, strongly reduce membrane binding affinity, attenuate membrane remodeling in vitro and alter subcellular localization. This effect is likely due to the loss of positive charge, which weakens interactions with negatively charged membranes. In Drosophila, acetylation-mimicking mutations of amphiphysin cause severe disruption of T-tubule organization and yield a flightless phenotype. Our data provide mechanistic insights into how lysine acetylation regulates membrane protein function, potentially impacting a plethora of membrane-related processes.

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The Abelson kinase and the Nedd4-family E3 ligases co-regulate Notch trafficking to limit signaling

Miranda-Alban,

Sanchez-Luege,

Valbuena

et al. 2024

Preprint

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Precise output from the conserved Notch signaling pathway governs a plethora of cellular processes and developmental transitions. Unlike other pathways that use a cytoplasmic relay, the Notch cell surface receptor transduces signaling directly to the nucleus, with endocytic trafficking providing critical regulatory nodes. Here we report that the cytoplasmic tyrosine kinase Abelson (Abl) facilitates Notch internalization into late endosomes/multivesicular bodies (LEs), thereby limiting signaling output in both ligand-dependent and -independent contexts. Abl phosphorylates the PPxY motif within Notch, a molecular target for its degradation via Nedd4-family ubiquitin ligases. We show that Su(dx), a family member, mediates the Abl-directed LE regulation of Notch via the PPxY, while another family member, Nedd4Lo, contributes to Notch internalization into LEs through both PPxY-dependent and independent mechanisms. Our findings demonstrate how a network of post-translational modifiers converging at LEs cooperatively modulate Notch signaling to ensure the precision and robustness of its cellular and developmental functions.

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DrosophilaNedd4-long reduces Amphiphysin levels in muscles and leads to impaired T-tubule formation

Abstract: An isoform of the fly ubiquitin ligase Nedd4 binds and degrades Amphiphysin, a postsynaptic and transverse tubule (T-tubule) protein in flies, thus impairing T-tubule formation and muscle function.

Cited by 5 publications

References 37 publications

Regulation of SH3PX1 by dNedd4-long at the Drosophila neuromuscular junction

Regulation of SH3PX1 by dNedd4-long at the Drosophila neuromuscular junction

Lysine acetylation regulates the interaction between proteins and membranes

The Abelson kinase and the Nedd4-family E3 ligases co-regulate Notch trafficking to limit signaling

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