Arf-like Protein 2 (ARL2) Controls Microtubule Neogenesis during Early Postnatal Photoreceptor Development

Arf-like protein 2 (ARL2) is a ubiquitously expressed small GTPase with multiple functions. In a cell culture, ARL2 participates with tubulin cofactor D (TBCD) in the neogenesis of tubulin αβ-heterodimers, the building blocks of microtubules. To evaluate this function in the retina, we conditionally deleted ARL2 in mouse retina at two distinct stages, either during the embryonic development (retArl2−/−) or after ciliogenesis specifically in rods (rodArl2−/−). retArl2−/− retina sections displayed distorted nuclear layers and a disrupted microtubule cytoskeleton (MTC) as early as postnatal day 6 (P6). Rod and cone outer segments (OS) did not form. By contrast, the rod ARL2 knockouts were stable at postnatal day 35 and revealed normal ERG responses. Cytoplasmic dynein is reduced in retArl2−/− inner segments (IS), suggesting that dynein may be unstable in the absence of a normal MTC. We investigated the microtubular stability in the absence of either ARL2 (retARL2−/−) or DYNC1H1 (retDync1h1−/−), the dynein heavy chain, and found that both the retArl2−/− and retDync1h1−/− retinas exhibited reduced microtubules and nuclear layer distortion. The results suggest that ARL2 and dynein depend on each other to generate a functional MTC during the early photoreceptor development.

[1]  K. Takemaru,et al.  Deletion of CEP164 in mouse photoreceptors post-ciliogenesis interrupts ciliary intraflagellar transport (IFT) , 2022, bioRxiv.

[2]  W. Baehr,et al.  Conditional Deletion of Cytoplasmic Dynein Heavy Chain in Postnatal Photoreceptors , 2021, Investigative ophthalmology & visual science.

[3]  W. Baehr,et al.  Review: Cytoplasmic dynein motors in photoreceptors , 2021, Molecular vision.

[4]  W. Baehr,et al.  Effect of conditional deletion of cytoplasmic dynein heavy chain DYNC1H1 on postnatal photoreceptors , 2021, PloS one.

[5]  U. Wolfrum,et al.  Roles for ELMOD2 and Rootletin in ciliogenesis , 2021, Molecular biology of the cell.

[6]  V. Arshavsky,et al.  Deletion of the phosphatase INPP5E in the murine retina impairs photoreceptor axoneme formation and prevents disc morphogenesis , 2021, The Journal of biological chemistry.

[7]  R. Kahn,et al.  The ARF GAP ELMOD2 acts with different GTPases to regulate centrosomal microtubule nucleation and cytokinesis , 2020, Molecular biology of the cell.

[8]  R. Kahn,et al.  ARF Family GTPases with Links to Cilia. , 2020, American journal of physiology. Cell physiology.

[9]  M. Magiera,et al.  The tubulin code and its role in controlling microtubule properties and functions , 2020, Nature Reviews Molecular Cell Biology.

[10]  X. Liu,et al.  Elmod3 knockout leads to progressive hearing loss and abnormalities in cochlear hair cell stereocilia. , 2019, Human molecular genetics.

[11]  C. Rivolta,et al.  Mutations in ARL2BP, a protein required for ciliary microtubule structure, cause syndromic male infertility in humans and mice , 2019, PLoS genetics.

[12]  W. Baehr,et al.  Insights into photoreceptor ciliogenesis revealed by animal models , 2019, Progress in Retinal and Eye Research.

[13]  J. Cherfils,et al.  ARF GTPases and their GEFs and GAPs: concepts and challenges , 2019, Molecular biology of the cell.

[14]  R. Kahn,et al.  ELMOD2 regulates mitochondrial fusion in a mitofusin-dependent manner, downstream of ARL2 , 2019, Molecular biology of the cell.

[15]  Zi-Bing Jin,et al.  Whole‐exome sequencing identified ARL2 as a novel candidate gene for MRCS (microcornea, rod‐cone dystrophy, cataract, and posterior staphyloma) syndrome , 2018, Clinical genetics.

[16]  Zachary C. Wright,et al.  ADP-Ribosylation Factor-Like 2 (ARL2) regulates cilia stability and development of outer segments in rod photoreceptor neurons , 2018, Scientific Reports.

[17]  Ratnesh K Singh,et al.  ARL2BP, a protein linked to retinitis pigmentosa, is needed for normal photoreceptor cilia doublets and outer segment structure , 2018, Molecular biology of the cell.

[18]  J. Liao,et al.  Super-resolution architecture of mammalian centriole distal appendages reveals distinct blade and matrix functional components , 2018, Nature Communications.

[19]  T. Kuner,et al.  STED nanoscopy of the centrosome linker reveals a CEP68-organized, periodic rootletin network anchored to a C-Nap1 ring at centrioles , 2018, Proceedings of the National Academy of Sciences.

[20]  P. Griffin,et al.  Nucleotide Binding to ARL2 in the TBCD∙ARL2∙β-Tubulin Complex Drives Conformational Changes in β-Tubulin. , 2017, Journal of molecular biology.

[21]  R. Kahn,et al.  The ARL2 GTPase regulates mitochondrial fusion from the intermembrane space , 2017, Cellular logistics.

[22]  S. Vacher,et al.  Alterations in the balance of tubulin glycylation and glutamylation in photoreceptors leads to retinal degeneration , 2017, Journal of Cell Science.

[23]  R. Kahn,et al.  A Trimer Consisting of the Tubulin-specific Chaperone D (TBCD), Regulatory GTPase ARL2, and β-Tubulin Is Required for Maintaining the Microtubule Network* , 2017, The Journal of Biological Chemistry.

[24]  A. Wittinghofer,et al.  Sorting of lipidated cargo by the Arl2/Arl3 system , 2016, Small GTPases.

[25]  R. Taneja,et al.  Arl2- and Msps-dependent microtubule growth governs asymmetric division , 2016, The Journal of cell biology.

[26]  K. Corbett,et al.  Tubulin cofactors and Arl2 are cage-like chaperones that regulate the soluble αβ-tubulin pool for microtubule dynamics , 2015, eLife.

[27]  R. Kahn,et al.  Characterization of Recombinant ELMOD (Cell Engulfment and Motility Domain) Proteins as GTPase-activating Proteins (GAPs) for ARF Family GTPases* , 2014, The Journal of Biological Chemistry.

[28]  D. Sharon,et al.  Mutations in ARL2BP, encoding ADP-ribosylation-factor-like 2 binding protein, cause autosomal-recessive retinitis pigmentosa. , 2013, American journal of human genetics.

[29]  N. Shen,et al.  MicroRNA-195 targets ADP-ribosylation factor-like protein 2 to induce apoptosis in human embryonic stem cell-derived neural progenitor cells , 2013, Cell Death and Disease.

[30]  I. Vetter,et al.  The interplay between RPGR, PDEδ and Arl2/3 regulate the ciliary targeting of farnesylated cargo , 2013, EMBO reports.

[31]  G. Tian,et al.  Tubulin-specific chaperones: components of a molecular machine that assembles the α/β heterodimer. , 2013, Methods in cell biology.

[32]  V. Sheffield,et al.  ARL13B, PDE6D, and CEP164 form a functional network for INPP5E ciliary targeting , 2012, Proceedings of the National Academy of Sciences.

[33]  I. Vetter,et al.  Structural basis for Arl3‐specific release of myristoylated ciliary cargo from UNC119 , 2012, The EMBO journal.

[34]  W E Moerner,et al.  STED microscopy with optimized labeling density reveals 9-fold arrangement of a centriole protein. , 2012, Biophysical journal.

[35]  Hui Zhao,et al.  The Rd8 mutation of the Crb1 gene is present in vendor lines of C57BL/6N mice and embryonic stem cells, and confounds ocular induced mutant phenotypes. , 2012, Investigative ophthalmology & visual science.

[36]  P. Bastiaens,et al.  Arl2-GTP and Arl3-GTP regulate a GDI-like transport system for farnesylated cargo. , 2011, Nature chemical biology.

[37]  G. Tian,et al.  Effect of TBCD and its regulatory interactor Arl2 on tubulin and microtubule integrity , 2010, Cytoskeleton.

[38]  R. Kahn,et al.  Cofactor D Functions as a Centrosomal Protein and Is Required for the Recruitment of the γ-Tubulin Ring Complex at Centrosomes and Organization of the Mitotic Spindle* , 2008, Journal of Biological Chemistry.

[39]  G. Germino,et al.  A critical developmental switch defines the kinetics of kidney cyst formation after loss of Pkd1 , 2007, Nature Medicine.

[40]  A. Spektor,et al.  Cep97 and CP110 Suppress a Cilia Assembly Program , 2007, Cell.

[41]  R. Kahn,et al.  ELMOD2 Is an Arl2 GTPase-activating Protein That Also Acts on Arfs* , 2007, Journal of Biological Chemistry.

[42]  Alfred Wittinghofer,et al.  GEFs and GAPs: Critical Elements in the Control of Small G Proteins , 2007, Cell.

[43]  Tiansen Li,et al.  Focus on molecules: rootletin. , 2006, Experimental eye research.

[44]  A. Marcus,et al.  Arl2 and Arl3 regulate different microtubule-dependent processes. , 2006, Molecular biology of the cell.

[45]  Y. Sauve,et al.  Rhodopsin‐iCre transgenic mouse line for Cre‐mediated rod‐specific gene targeting , 2005, Genesis.

[46]  R. Kahn,et al.  Assays used in the analysis of Arl2 and its binding partners. , 2005, Methods in enzymology.

[47]  W. G. Kelly,et al.  Functional genomic analysis of the ADP‐ribosylation factor family of GTPases: phylogeny among diverse eukaryotes and function in C. elegans , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[48]  A. Wittinghofer,et al.  The complex of Arl2‐GTP and PDEδ: from structure to function , 2002 .

[49]  Min Han,et al.  The C. elegans evl-20 gene is a homolog of the small GTPase ARL2 and regulates cytoskeleton dynamics during cytokinesis and morphogenesis. , 2002, Developmental cell.

[50]  A. Wittinghofer,et al.  The complex of Arl2-GTP and PDE delta: from structure to function. , 2002, The EMBO journal.

[51]  D. Meinke,et al.  The TITAN5 Gene of Arabidopsis Encodes a Protein Related to the ADP Ribosylation Factor Family of GTP Binding Proteins , 2000, Plant Cell.

[52]  Nicholas J. Cowan,et al.  Adp Ribosylation Factor-like Protein 2 (Arl2) Regulates the Interaction of Tubulin-Folding Cofactor D with Native Tubulin , 2000, The Journal of cell biology.

[53]  B. Hogan,et al.  Retina‐ and ventral forebrain‐specific Cre recombinase activity in transgenic mice , 2000, Genesis.

[54]  R. Kahn,et al.  The ARF-like 2 (ARL2)-binding Protein, BART , 1999, The Journal of Biological Chemistry.

[55]  Nicholas J. Cowan,et al.  The α- and β-tubulin folding pathways , 1997 .

[56]  Christophe Ampe,et al.  Pathway Leading to Correctly Folded β-Tubulin , 1996, Cell.

[57]  R. Kahn,et al.  ADP-ribosylation factor (ARF)-like 3, a new member of the ARF family of GTP-binding proteins cloned from human and rat tissues. , 1994, The Journal of biological chemistry.

[58]  A. Schürmann,et al.  Cloning of two novel ADP-ribosylation factor-like proteins and characterization of their differential expression in 3T3-L1 cells. , 1994, The Journal of biological chemistry.

[59]  K. Kamata,et al.  Stabilization of microtubules by dynein-binding in vitro. Stability of microtubule-dynein complex. , 1993, Biochimica et biophysica acta.

[60]  Michael B. Yaffe,et al.  TCP1 complex is a molecular chaperone in tubulin biogenesis , 1992, Nature.

[61]  M. Scott,et al.  The arflike gene encodes an essential GTP-binding protein in Drosophila. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[62]  D. Botstein,et al.  Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes. , 1990, Molecular and cellular biology.