Interactome analysis reveals that FAM161A, deficient in recessive retinitis pigmentosa, is a component of the Golgi-centrosomal network.
Defects in FAM161A, a protein of unknown function localized at the cilium of retinal photoreceptor cells, cause retinitis pigmentosa, a form of hereditary blindness. By using different fragments of this protein as baits to screen cDNA libraries of human and bovine retinas, we defined a yeast two-hybrid-based FAM161A interactome, identifying 53 bona fide partners. In addition to statistically significant enrichment in ciliary proteins, as expected, this interactome revealed a substantial bias towards proteins from the Golgi apparatus, the centrosome and the microtubule network. Validation of interaction with key partners by co-immunoprecipitation and proximity ligation assay confirmed that FAM161A is a member of the recently recognized Golgi-centrosomal interactome, a network of proteins interconnecting Golgi maintenance, intracellular transport and centrosome organization. Notable FAM161A interactors included AKAP9, FIP3, GOLGA3, KIFC3, KLC2, PDE4DIP, NIN and TRIP11. Furthermore, analysis of FAM161A localization during the cell cycle revealed that this protein followed the centrosome during all stages of mitosis, likely reflecting a specific compartmentalization related to its role at the ciliary basal body during the G0 phase. Altogether, these findings suggest that FAM161A's activities are probably not limited to ciliary tasks but also extend to more general cellular functions, highlighting possible novel mechanisms for the molecular pathology of retinal disease.
Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland.
Silvio Alessandro
SA
Di Gioia
Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland.
Pietro
P
Farinelli
Department of Human Genetics and Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands and.
Stef J F
SJ
Letteboer
Unit of Gene Therapy and Stem Cell Biology, Jules-Gonin Eye Hospital, University of Lausanne, Lausanne, Switzerland.
Yvan
Y
Arsenijevic
Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Dror
D
Sharon
Department of Human Genetics and Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands and.
Ronald
R
Roepman
Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland carlo.rivolta@unil.ch.
Carlo
C
Rivolta
D000818:Animals; D002352:Carrier Proteins; D002417:Cattle; D002460:Cell Line; D018385:Centrosome / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism*; D005808:Genes, Recessive; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps; D021381:Protein Transport; D012174:Retinitis Pigmentosa / Q000235:genetics* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D002352:Carrier Proteins; D005136:Eye Proteins; C552553:FAM161A protein, human