Tubby-like protein 1 (TULP1) interacts with F-actin in photoreceptor cells.
OBJECTIVE
TULP1 is a photoreceptor-specific protein of unknown function that, when mutated, can cause retinitis pigmentosa in humans and photoreceptor degeneration in mice. Toward a better understanding of the role of TULP1 in retinal disease, its subcellular localization was sought and the TULP1 protein binding partners identified.


METHODS
Immunocytochemistry and subcellular fractionation were used to determine the localization of TULP1 and actin in COS7 cells and photoreceptor cells. Immunoprecipitation from retinal lysates followed by liquid chromatography tandem mass spectrometry and in vitro binding assays was used to identify TULP1-binding partners. Phospholipid binding assays were performed with a commercially available kit.


RESULTS
TULP1 localizes at or near the plasma membrane and associates with the membranous fraction of COS7 cells, probably through binding phosphorylated phospholipids. In addition, TULP1 partitions to the aqueous phase during Triton X-114 extraction. Immunoprecipitation from retinal lysate identified F-actin as a possible TULP1-binding partner. Co-sedimentation assays further support an interaction between TULP1 and actin. In photoreceptor cells, actin and TULP1 colocalize at the inner segment, connecting cilium, and outer limiting membrane.


CONCLUSIONS
TULP1 is a cytoplasmic protein that associates with cellular membranes and the cytoskeleton. TULP1 and actin appear to interact and colocalize in photoreceptor cells of the retina. TULP1 may be involved in actin cytoskeletal functions such as protein trafficking that takes place at or near the plasma membrane from the inner segment through the connecting cilium into the outer segment of photoreceptor cells.
Xi
Quansheng
Q
Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Pauer
Gayle J T
GJ
Marmorstein
Alan D
AD
Crabb
John W
JW
Hagstrom
Stephanie A
SA
D000199:Actins / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002417:Cattle; D002522:Chlorocebus aethiops; D002853:Chromatography, Liquid; D005136:Eye Proteins / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D008401:Gas Chromatography-Mass Spectrometry; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D013347:Subcellular Fractions; D014162:Transfection
D000199:Actins; D005136:Eye Proteins; C105443:TULP1 protein, human