Subcellular localisation of recombinant alpha- and gamma-synuclein.
alpha-Synuclein, a protein implicated in neurodegenerative diseases and of elusive physiological function owes its name to an observed presence in presynaptic and nuclear compartments. However, its nuclear localisation has remained controversial. We expressed synuclein-eGFP fusion proteins in organotypic rat hippocampal slice cultures and murine hippocampal primary neurons using a Sindbis virus expression system. Recombinant full-length alpha-synuclein accumulated in presynaptic locations, mimicking its native distribution. Expression of deletion mutant alpha-synuclein revealed that presynaptic targeting depended on the presence of its N-terminal and core region. This domain also causes nuclear exclusion of the alpha-synuclein fusion protein. In contrast, the C-terminal domain of alpha-synuclein directs fusion proteins into the nuclear compartment. The related protein gamma-synuclein contains a similar N-terminal and core domain as alpha-synuclein. However, gamma-synuclein lacks a C-terminal domain that causes nuclear localisation of the fusion protein, suggesting that the two synucleins might have different roles relating to the cell nucleus.
Specht
Christian G
CG
Laboratory for Molecular Pharmacology, Department of Pharmacology, University College London, UCL, London WC1E 6BT, UK.
Tigaret
Cezar M
CM
Rast
Georg F
GF
Thalhammer
Agnes
A
Rudhard
York
Y
Schoepfer
Ralf
R
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D002451:Cell Compartmentation / Q000502:physiology; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009154:Mutation / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000378:metabolism* / Q000648:ultrastructure; D017434:Protein Structure, Tertiary / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D051843:Synucleins; D051844:alpha-Synuclein; D051845:gamma-Synuclein
D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; C497605:Snca protein, mouse; C497606:Snca protein, rat; D051843:Synucleins; D051844:alpha-Synuclein; D051845:gamma-Synuclein