A collection of caged compounds for probing roles of local translation in neurobiology.
Spatially localized translation plays a vital role in the normal functioning of neuronal systems and is widely believed to be involved in both learning and memory formation. It is of central interest to understand both the phenomenon and molecular mechanisms of local translation using new tools and approaches. Caged compounds can, in principle, be used as tools to investigate local translation since optical activation of bioactive molecules can achieve both spatial and temporal resolution on the micron scale and on the order of seconds or less, respectively. Successful caging of bioactive molecules requires the identification of key functional groups in appropriate molecules and the introduction of a suitable caging moiety. Here we present the design, synthesis and testing of a collection of three caged compounds: anisomycin caged with a diethylaminocoumarin moiety and dimethoxynitrobenzyl caged versions of 4E-BP and rapamycin. Whereas caged anisomycin can be used to control general translation, caged 4E-BP serves as a probe of cap-dependent translation initiation and caged rapamycin serves a probe of the role of mTORC1 in translation initiation. In vitro translation assays demonstrate that these caging strategies, in combination with the aforementioned compounds, are effective for optical control making it likely that such strategies can successfully employed in the study of local translation in living systems.
Sadovski
Oleg
O
Department of Chemistry, University of Toronto, 80 St. George ST., Toronto, ON, Canada M5S 3H6.
Jaikaran
Anna S I
AS
Samanta
Subhas
S
Fabian
Marc R
MR
Dowling
Ryan J O
RJ
Sonenberg
Nahum
N
Woolley
G Andrew
GA
D049933:Aminocoumarins / Q000737:chemistry*; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D000076222:Mechanistic Target of Rapamycin Complex 1; D046912:Multiprotein Complexes; D009420:Nervous System / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D011817:Rabbits; D020123:Sirolimus / Q000494:pharmacology; D058570:TOR Serine-Threonine Kinases
D049933:Aminocoumarins; D046912:Multiprotein Complexes; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D012313:RNA; D000841:Anisomycin; D000076222:Mechanistic Target of Rapamycin Complex 1; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus