Arl4D-EB1 interaction promotes centrosomal recruitment of EB1 and microtubule growth

Abstract

ADP-ribosylation factor (Arf)-like 4D (Arl4D), one of the Arf-like small GTPases, functions in the regulation of cell morphology, cell migration, and actin cytoskeleton remodeling. EB1 is a microtubule plus-end tracking protein that preferentially localizes at the tips of the plus ends of growing microtubules and at the centrosome. EB1 depletion results in many centrosome-related defects. Here, we report that Arl4D promotes the recruitment of EB1 to the centrosome and regulates microtubule nucleation. We first showed that Arl4D interacts with EB1 in a GTP-dependent manner. This interaction is dependent on the C-terminal EBH region of EB1 and partially dependent on an SxLP motif of Arl4D. We found that Arl4D colocalized with γ-tubulin in centrosomes and the depletion of Arl4D resulted in a centrosomal microtubule nucleation defect. We further demonstrated that abolishing Arl4D-EB1 interaction decreased microtubule nucleation rate and diminished the centrosomal recruitment of EB1 without affecting MT growth rate. In addition, Arl4D binding to EB1 increased the association between the p150 subunit of dynactin and EB1, which is important for microtubule stabilization. Together, our results indicate that Arl4D modulates microtubule nucleation through regulation of the EB1-p150 association at the centrosome. [Media: see text]

Identification and validation of seven RNA binding protein genes as a prognostic signature in oral cavity squamous cell carcinoma

“…ADP-ribosylation factor (Arf)-like 4D (ARL4D) is an Arf-like small GTPase that regulates actin cytoskeleton remodeling, cell morphology and migration [ 24 ]. Lin et al reported that ARL4D plays an important role in microtubule growth [ 25 ]. Upregulated ARL4D promotes neurite outgrowth in N1E-115 neuroblastoma cells [ 26 ].…”

Mapping the global interactome of the ARF family reveals spatial organization in cellular signaling pathways

“…Pathway enrichment analyses of individual clusters provide previously unknown localizations and functions for several ARFs. Cluster 5 is enriched in preys specifically identified with ARL4D, a GTPase known for its roles in actin remodeling (Li et al, 2007), neurite formation (Yamauchi et al, 2009), adipogenesis (Yu et al, 2011), microtubule growth (Lin et al, 2020), and immune functions (Geers et al, 2022) (Tolksdorf et al, 2018). Overrepresentation analyses of Gene ontology biological processes (GO BP) are consistent with ARL4D residency in the nucleus and potential functions in the nucleolus, but also reveals roles in ribosome biogenesis, regulation of RNA splicing, regulation of transcription, nucleosome organization, regulation of macromolecules biosynthetic process and RNA processing and modification (Fig.…”

“…Numerous other ARLs, including ARL2/3, ARL4D, ARL6, and ARL13B, play regulatory function in microtubule-associated processes within centrosomes, spindles, midbodies, basal bodies and cilia (Yong et al, 2020, Zhou et al, 2006, Bhattarai et al, 2019, Lin et al, 2020, Nachury et al, 2007, Li et al, 2010. The heterogeneous spatial distribution of some ARL proteins can drive interplay with multiple cellular processes.…”

“…Notably, ARL8A and ARL8B localize to lysosomes, facilitating their motility (Hofmann and Munro, 2006). Numerous other ARLs, including ARL2/3, ARL4D, ARL6, and ARL13B, play regulatory function in microtubule-associated processes within centrosomes, spindles, midbodies, basal bodies and cilia (Yong et al, 2020, Zhou et al, 2006, Bhattarai et al, 2019, Lin et al, 2020, Nachury et al, 2007, Li et al, 2010). The heterogeneous spatial distribution of some ARL proteins can drive interplay with multiple cellular processes.…”

The ADP-Ribosylation Factor 4d Restricts Regulatory T-Cell Induction via Control of IL-2 Availability

“…The ADP ribosylation factor 4d is part of the large family of ARF GTPases, mainly known for their role in membrane transport processes [ 9 ]. A functional role for Arl4d itself has been shown in actin remodeling [ 10 ], adipocyte development [ 11 ], neurite outgrowth [ 12 ], and microtubule growth [ 13 ], and Arl4d has known interaction partners (the cytohesin protein family) with immune function [ 14 , 15 ]. We could show that Arl4d expression is induced in CD8 T cells during priming by liver sinusoidal endothelial cells (LSEC), which is dependent on PD-L1/PD-1 signaling.…”