Single molecule analysis reveals reversible and irreversible steps during spliceosome activation.

Abstract:

:The spliceosome is a complex machine composed of small nuclear ribonucleoproteins (snRNPs) and accessory proteins that excises introns from pre-mRNAs. After assembly the spliceosome is activated for catalysis by rearrangement of subunits to form an active site. How this rearrangement is coordinated is not well-understood. During activation, U4 must be released to allow U6 conformational change, while Prp19 complex (NTC) recruitment is essential for stabilizing the active site. We used multi-wavelength colocalization single molecule spectroscopy to directly observe the key events in Saccharomyces cerevisiae spliceosome activation. Following binding of the U4/U6.U5 tri-snRNP, the spliceosome either reverses assembly by discarding tri-snRNP or proceeds to activation by irreversible U4 loss. The major pathway for NTC recruitment occurs after U4 release. ATP stimulates both the competing U4 release and tri-snRNP discard processes. The data reveal the activation mechanism and show that overall splicing efficiency may be maintained through repeated rounds of disassembly and tri-snRNP reassociation.

journal_name

Elife

journal_title

eLife

authors

Hoskins AA,Rodgers ML,Friedman LJ,Gelles J,Moore MJ

doi

10.7554/eLife.14166

subject

Has Abstract

pub_date

2016-05-31 00:00:00

issn

2050-084X

journal_volume

5

pub_type

杂志文章

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