Differential effects of changes in the length of a signal anchor domain on membrane insertion, subunit assembly, and intracellular transport of a type II integral membrane protein

被引:8
作者
Parks, GD
机构
[1] Dept. of Microbiology and Immunology, Wake Forest Univ. Medical Center, Winston-Salem
[2] Dept. of Microbiology and Immunology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27157-1064, Medical Center Blvd.
关键词
D O I
10.1074/jbc.271.12.7187
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The length requirement for a functional uncleaved signal/anchor (S/A) domain of the paramyxovirus hemagglutinin-neuraminidase (HN) type II glycoprotein was analyzed. HN mutants with progressive NH2-terminal S/A deletions or insertions were expressed in HeLa cells, and the membrane targeting, folding, tetramer assembly, and intracellular transport of the proteins were examined. Changing the length of the S/A by two residues resulted in HN mutants that displayed aberrant endoplasmic reticulum (ER) membrane targeting or translocation. This phenotype did not simply reflect upper or lower limitations on the size of a functional S/A, because normal signaling was restored by further alterations involving three or four residues. Likewise, ER-to-Golgi transport of mutants containing deletions of one or two S/A residues was delayed (similar to 30% of WT) or blocked, but transport was restored for a mutant with a total of three deleted residues. HN mutants with S/A insertions of three or four Leu residues differed from wild-type HN by having heterogeneous Golgi-specific carbohydrate modifications. Differences in ER-to-Golgi transport of the mutants did not strictly correlate with defects in either native folding of the ectodomain or the assembly of two dimers into a tetramer. Together, these data suggest that efficient entry into and exit from the ER are sensitive to changes in the HN S/A that may reflect alterations to a structural requirement along one side of an alpha-helix.
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页码:7187 / 7195
页数:9
相关论文
共 52 条
[1]   STRUCTURAL REQUIREMENTS OF A MEMBRANE-SPANNING DOMAIN FOR PROTEIN ANCHORING AND CELL-SURFACE TRANSPORT [J].
ADAMS, GA ;
ROSE, JK .
CELL, 1985, 41 (03) :1007-1015
[2]  
ASHFORD DA, 1993, J BIOL CHEM, V268, P3260
[3]   NH2-TERMINAL HYDROPHOBIC REGION OF INFLUENZA-VIRUS NEURAMINIDASE PROVIDES THE SIGNAL FUNCTION IN TRANSLOCATION [J].
BOS, TJ ;
DAVIS, AR ;
NAYAK, DP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (08) :2327-2331
[4]   CHOLESTEROL AND THE GOLGI-APPARATUS [J].
BRETSCHER, MS ;
MUNRO, S .
SCIENCE, 1993, 261 (5126) :1280-1281
[5]   REDUNDANCY OF SIGNAL AND ANCHOR FUNCTIONS IN THE NH2-TERMINAL UNCHARGED REGION OF INFLUENZA-VIRUS NEURAMINIDASE, A CLASS-II MEMBRANE GLYCOPROTEIN [J].
BROWN, DJ ;
HOGUE, BG ;
NAYAK, DP .
JOURNAL OF VIROLOGY, 1988, 62 (10) :3824-3831
[6]   AN AUTONOMOUS SIGNAL FOR BASOLATERAL SORTING IN THE CYTOPLASMIC DOMAIN OF THE POLYMERIC IMMUNOGLOBULIN RECEPTOR [J].
CASANOVA, JE ;
APODACA, G ;
MOSTOV, KE .
CELL, 1991, 66 (01) :65-75
[7]   VAL-]ALA MUTATIONS SELECTIVELY ALTER HELIX-HELIX PACKING IN THE TRANSMEMBRANE SEGMENT OF PHAGE M13 COAT PROTEIN [J].
DEBER, CM ;
KHAN, AR ;
LI, ZM ;
JOENSSON, C ;
GLIBOWICKA, M ;
WANG, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (24) :11648-11652
[8]   FOLDING AND ASSEMBLY OF VIRAL MEMBRANE-PROTEINS [J].
DOMS, RW ;
LAMB, RA ;
ROSE, JK ;
HELENIUS, A .
VIROLOGY, 1993, 193 (02) :545-562
[9]  
ENGLEMAN DM, 1986, ANN REV BIOPHYS CHEM, V15, P321
[10]   EUKARYOTIC TRANSIENT-EXPRESSION SYSTEM BASED ON RECOMBINANT VACCINIA VIRUS THAT SYNTHESIZES BACTERIOPHAGE-T7 RNA-POLYMERASE [J].
FUERST, TR ;
NILES, EG ;
STUDIER, FW ;
MOSS, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (21) :8122-8126