The upstream 5′ splice site remains associated to the transcription machinery during intron synthesis

Yodfat Leader, Galit Lev Maor (), Matan Sorek, Ronna Shayevitch, Maram Hussein, Ofir Hameiri, Luna Tammer, Jonathan Zonszain, Ifat Keydar, Dror Hollander, Eran Meshorer and Gil Ast ()
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Yodfat Leader: Tel-Aviv University
Galit Lev Maor: Tel-Aviv University
Matan Sorek: The Hebrew University of Jerusalem, Edmond J. Safra Campus
Ronna Shayevitch: Tel-Aviv University
Maram Hussein: Tel-Aviv University
Ofir Hameiri: Tel-Aviv University
Luna Tammer: Tel-Aviv University
Jonathan Zonszain: Tel-Aviv University
Ifat Keydar: Tel-Aviv University
Dror Hollander: Tel-Aviv University
Eran Meshorer: The Hebrew University of Jerusalem, Edmond J. Safra Campus
Gil Ast: Tel-Aviv University

Nature Communications, 2021, vol. 12, issue 1, 1-11

Abstract: Abstract In the earliest step of spliceosome assembly, the two splice sites flanking an intron are brought into proximity by U1 snRNP and U2AF along with other proteins. The mechanism that facilitates this intron looping is poorly understood. Using a CRISPR interference-based approach to halt RNA polymerase II transcription in the middle of introns in human cells, we discovered that the nascent 5′ splice site base pairs with a U1 snRNA that is tethered to RNA polymerase II during intron synthesis. This association functionally corresponds with splicing outcome, involves bona fide 5′ splice sites and cryptic intronic sites, and occurs transcriptome-wide. Overall, our findings reveal that the upstream 5′ splice sites remain attached to the transcriptional machinery during intron synthesis and are thus brought into proximity of the 3′ splice sites; potentially mediating the rapid splicing of long introns.

Date: 2021
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DOI: 10.1038/s41467-021-24774-6

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