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. Author manuscript; available in PMC: 2012 Nov 28.
Published in final edited form as: Trends Biochem Sci. 2012 Apr 3;37(5):179–188. doi: 10.1016/j.tibs.2012.02.009

Figure 4.

Figure 4

Schematic for single molecule fluorescence analysis of splicing. (a) Hoskins et al. tethered fluorescent pre-mRNAs labeled with Alexa488 to a glass surface and then monitored how fluorescent spliceosome subcomplexes (e.g. U2-SF3 and U4/U6.U5 tri-snRNP) associated with the pre-mRNA. Fluctuations in fluorescence intensity indicated that the subcomplexes can associate multiple times with a given pre-mRNA. (b) By incorporating a FRET donor (Cy3) and acceptor (Cy5) pair into a surface-tethered pre-mRNA, Abelson et al. were able to show that pre-mRNA conformation reversibly fluctuated during splicing. These conformational changes are shown by changes in the FRET signal of single pre-mRNA molecules. Data in (a) and (b) are from references [45] and [49], respectively, and used with the authors’ permission.

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