A LIN28-dependent structural change in pre-let-7g directly inhibits dicer processing.

Abstract:

:Several recent studies have provided evidence that LIN28, a cytoplasmic RNA-binding protein, inhibits the biogenesis of members of the let-7 microRNA family at the Dicer step in both mammals and Caenorhabditis elegans. However, the precise mechanism of inhibition is still poorly understood. Here we report on an in vitro study, which combined RNase footprinting, gel shift binding assays, and processing assays, to investigate the molecular basis and function of the interaction between the native let-7g precursor (pre-let-7g) and LIN28. We have mapped the structure of pre-let-7g and identified some regions of the terminal loop of pre-let-7g that physically interact with LIN28. We have also identified a conformational change upon LIN28 binding that results in the unwinding of an otherwise double-stranded region at the Dicer processing site of pre-let-7g. Furthermore, we showed that a mutant pre-let-7g that displays an open upper stem inhibited pre-let-7g Dicer processing to the same extent as LIN28. The data support a mechanism by which LIN28 can directly inhibit let-7g biogenesis at the Dicer processing step.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Lightfoot HL,Bugaut A,Armisen J,Lehrbach NJ,Miska EA,Balasubramanian S

doi

10.1021/bi200851d

subject

Has Abstract

pub_date

2011-09-06 00:00:00

pages

7514-21

issue

35

eissn

0006-2960

issn

1520-4995

journal_volume

50

pub_type

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