Identification of an outer segment targeting signal in the COOH terminus of rhodopsin using transgenic Xenopus laevis

被引:160
作者
Tam, BM [1 ]
Moritz, OL [1 ]
Hurd, LB [1 ]
Papermaster, DS [1 ]
机构
[1] Univ Connecticut, Ctr Hlth, Dept Pharmacol, Program Neurosci, Farmington, CT 06030 USA
关键词
rhodopsin; photoreceptors; transgenic animals; membrane proteins; cell polarity;
D O I
10.1083/jcb.151.7.1369
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Mislocalization of the photopigment rhodopsin may be involved in the pathology of certain inherited retinal degenerative diseases. Here, we have elucidated rhodopsin's targeting signal which is responsible for its polarized distribution to the rod outer segment (ROS). Various green fluorescent protein (GFP)/rhodopsin COOH-terminal fusion proteins were expressed specifically in the major red rod photoreceptors of transgenic Xenopus laevis under the control of the Xenopus opsin promoter. The fusion proteins were targeted to membranes via lipid modifications (palmitoylation and myristoylation) as opposed to membrane spanning domains. Membrane association was found to be necessary but not sufficient for efficient ROS localization. A GFP fusion protein containing only the cytoplasmic COOH-terminal 44 amino acids of Xenopus rhodopsin localized exclusively to ROS membranes. Chimeras between rhodopsin and oc adrenergic receptor COOH-terminal sequences further refined rhodopsin's ROS localization signal to its distal eight amino acids. Mutations/deletions of this region resulted in partial delocalization of the fusion proteins to rod inner segment (RIS) membranes. The targeting and transport of endogenous wildtype rhodopsin was unaffected by the presence of mislocalized GFP fusion proteins.
引用
收藏
页码:1369 / 1380
页数:12
相关论文
共 74 条
[1]   O-linked glycans mediate apical sorting of human intestinal sucrase-isomaltase through association with lipid rafts [J].
Alfalah, M ;
Jacob, R ;
Preuss, U ;
Zimmer, KP ;
Naim, H ;
Naim, HY .
CURRENT BIOLOGY, 1999, 9 (11) :593-596
[2]   Multiple sorting signals determine apical localization of a nonglycosylated integral membrane protein [J].
Alonso, MA ;
Fan, L ;
Alarcón, B .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (49) :30748-30752
[3]  
Amaya E, 1999, METH MOL B, V97, P393, DOI 10.1385/1-59259-270-8:393
[4]   Characterization of the Xenopus rhodopsin gene [J].
Batni, S ;
Scalzetti, L ;
Moody, SA ;
Knox, BE .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (06) :3179-3186
[5]   The basolateral localization signal of the follicle-stimulating hormone receptor [J].
Beau, I ;
Groyer-Picard, MT ;
Le Bivic, A ;
Vannier, B ;
Loosfelt, H ;
Milgrom, E ;
Misrahi, M .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (29) :18610-18616
[6]   IMMUNOCYTOCHEMICAL LOCALIZATION OF OPSIN IN ROD PHOTORECEPTORS DURING PERIODS OF RAPID DISC ASSEMBLY [J].
BESHARSE, JC ;
WETZEL, MG .
JOURNAL OF NEUROCYTOLOGY, 1995, 24 (05) :371-388
[7]   MEMBRANE ASSEMBLY IN RETINAL PHOTORECEPTORS .1. FREEZE-FRACTURE ANALYSIS OF CYTOPLASMIC VESICLES IN RELATIONSHIP TO DISK ASSEMBLY [J].
BESHARSE, JC ;
PFENNINGER, KH .
JOURNAL OF CELL BIOLOGY, 1980, 87 (02) :451-463
[8]   MECHANISM OF MEMBRANE ANCHORING AFFECTS POLARIZED EXPRESSION OF 2 PROTEINS IN MDCK CELLS [J].
BROWN, DA ;
CRISE, B ;
ROSE, JK .
SCIENCE, 1989, 245 (4925) :1499-1501
[9]   RHODOPSIN ROTATES IN VISUAL RECEPTOR MEMBRANE [J].
BROWN, PK .
NATURE-NEW BIOLOGY, 1972, 236 (63) :35-&
[10]   COLOCALIZATION OF SYNAPTOPHYSIN WITH TRANSFERRIN RECEPTORS - IMPLICATIONS FOR SYNAPTIC VESICLE BIOGENESIS [J].
CAMERON, PL ;
SUDHOF, TC ;
JAHN, R ;
DECAMILLI, P .
JOURNAL OF CELL BIOLOGY, 1991, 115 (01) :151-164