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Contributo in rivista
Tipo: Articolo in rivista
Titolo: ERK1 nucleocytoplasmic shuttling rate depends on specific N-terminal aminoacids.
Anno di pubblicazione: 2010
Autori: Marchi M, Pancrazi L, Maffei M, Ratto GM, Costa M.
Affiliazioni autori: 1. Univ Hosp Pisa, Dept Endocrinol & Kidney, Dulbecco Telethon Inst, Pisa, Italy (Marchi M., Pancrazi L., Maffei M.) 2. Scuola Normale Super Pisa, NEST INFM, Pisa, Italy (Marchi M., Ratto G.M.) 3. CNR, Inst Food Sci, Avellino, Italy (Pancrazi L., Maffei M.)
Abstract: Despite ERK1 and ERK2 were considered interchangeable isoforms for a long time, their roles are now emerging as only partially overlapping. We recently reported that the nucleocytoplasmic trafficking of GFP-tagged ERK1 is slower than that of ERK2, this difference being caused by a unique domain of ERK1 located at its N-terminus (ERK1-Nt). In the present report we further investigated this issue by asking which were the specific aminoacids involved in such process. By photobleaching strategy, we demonstrated that ERK1-Nt is a domain capable to slow down the nucleocytoplasmic shuttling rate even of a small cargo protein. ERK1-Nt was then dissected into three regions as follows: 1 (aa 1-9), 2 (aa 10-29) and 3, (aa 30-39) that were deleted or mutated at specific sites. Dynamic imaging assessment of the role played by each region in determining the shuttling rate revealed that: region 1 has no significant role, region 2 and specific aminoacids of region 3 (V-31, K-33, P-36) are critical, but singularly do not totally account for the difference in the shuttling rate between ERK1 and 2. Finally, we demonstrated that the nucleocytoplasmic shuttling rate of a passively diffusing protein (mRED) is inversely related to ERK1-Nt-GFP concentrations inside the cell, thus suggesting that ERK1-Nt-GFP occupies the nuclear pore perhaps because of an important affinity of ERK1-Nt for nucleoporins. In conclusion, ERK1-Nt is a domain able per se to confer a slower shuttling rate to a cargo protein. Specific regions within this domain were identified as responsible for this biophysical property. (C) 2010 Elsevier Inc. All rights reserved.
Pagine da: 166
Pagine a: 172
Biochemical and biophysical research communications
Numero volume: 398
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