Anterograde and retrograde intracellular trafficking of fluorescent cellular prion protein

被引:40
作者
Hachiya, NS
Watanabe, K
Yamada, M
Sakasegawa, Y
Kaneko, K [1 ]
机构
[1] Japan Sci & Technol Agcy, NCNP, NIN, Dept Cortical Funct Disorders, Tokyo 1878502, Japan
[2] Japan Sci & Technol Agcy, CREST, Tokyo 1878502, Japan
关键词
cellular prion protein; green fluorescent protein; microtubules; kinesin family; dynein;
D O I
10.1016/j.bbrc.2004.01.126
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
In order to investigate the microtubule-associated intracellular trafficking of the NH2-terminal cellular prion protein (PrPC) fragment [Biochem. Biophys. Res. Commun. 313 (2004) 818], we performed a real-time imaging of fluorescent PrPC (GFP-PrPC) in living cells. Such GFP-PrPC exhibited an anterograde movement towards the direction of plasma membranes at a speed of 140180 nm/s, and a retrograde movement inwardly at a speed of 1.0-1.2 mum,/s. The anterograde and retrograde movements of GFP-PrPC were blocked by a kinesin family inhibitor (AMP-PNP) and a dynein family inhibitor (vanadate), respectively. Furthermore, anti-kinesin antibody (alpha-kinesin) blocked its anterograde motility, whereas anti-dynein antibody (alpha-dynein) blocked its retrograde motility. These data suggested the kinesin family-driven anterograde and the dynein-driven retrograde movements of GFP-PrPC. Mapping of the interacting domains of PrPC identified amino acid residues indispensable for interactions with kinesin family: NH2-terminal mouse (Mo) residues 53-91 and dynein: NH2-terminal Mo residues 23-33, respectively. Our findings argue that the discrete N-terminal amino acid residues are indispensable for the anterograde and retrograde intracellular movements of PrPC. (C) 2004 Elsevier Inc. All rights reserved.
引用
收藏
页码:802 / 807
页数:6
相关论文
共 36 条
[1]  
Bamborough P, 1996, COLD SPRING HARB SYM, V61, P495
[2]   Spongiform encephalopathies - B lymphocytes and neuroinvasion [J].
Brown, P .
NATURE, 1997, 390 (6661) :662-663
[3]   SCRAPIE-INFECTED MURINE NEURO-BLASTOMA CELLS PRODUCE PROTEASE-RESISTANT PRION PROTEINS [J].
BUTLER, DA ;
SCOTT, MRD ;
BOCKMAN, JM ;
BORCHELT, DR ;
TARABOULOS, A ;
HSIAO, KK ;
KINGSBURY, DT ;
PRUSINER, SB .
JOURNAL OF VIROLOGY, 1988, 62 (05) :1558-1564
[4]   Copper binding to the octarepeats of the prion protein - Affinity, specificity, folding, and cooperativity: Insights from circular dichroism [J].
Garnett, AP ;
Viles, JH .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (09) :6795-6802
[5]   Microtubules-associated intracellular localization of the NH2-terminal cellular prion protein fragment [J].
Hachiya, NS ;
Watanabe, K ;
Sakasegawa, Y ;
Kaneko, K .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 313 (03) :818-823
[6]   Kinesin and dynein superfamily proteins and the mechanism of organelle transport [J].
Hirokawa, N .
SCIENCE, 1998, 279 (5350) :519-526
[7]   Dynamics and mechanics of the microtubule plus end [J].
Howard, J ;
Hyman, AA .
NATURE, 2003, 422 (6933) :753-758
[8]   Mutant prion proteins axe partially retained in the endoplasmic reticulum [J].
Ivanova, L ;
Barmada, S ;
Kummer, T ;
Harris, DA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (45) :42409-42421
[9]   Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation [J].
Kaneko, K ;
Zulianello, L ;
Scott, M ;
Cooper, CM ;
Wallace, AC ;
James, TL ;
Cohen, FE ;
Prusiner, SB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (19) :10069-10074
[10]   Prion protein binds copper within the physiological concentration range [J].
Kramer, ML ;
Kratzin, HD ;
Schmidt, B ;
Römer, A ;
Windl, O ;
Liemann, S ;
Hornemann, S ;
Kretzschmar, H .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (20) :16711-16719