Insights into the structure and evolution of the human SAGA complex by affinity-ligand purification

TL;DR

This study reveals the structure of the human SAGA complex and how it incorporates a splicing module unique to metazoans.

Contribution

The paper presents the high-resolution structure of the SPL module and its integration into SAGA using unperturbed human cells.

Findings

The SPL module and TAF6L HEAT repeat domain structure were resolved at high resolution.

TAF6L and TAF5L have structural differences that allow SPL integration into SAGA.

SPL interacts with SAGA through a smaller interface than in U2snRNP, suggesting a potential relay to splicing machinery.

Abstract

Human SAGA is a 20-subunit transcriptional coactivator. Compared with yeast, metazoan SAGA uniquely incorporates a 150-kDa splicing-factor module (SPL), also present in U2 small nuclear ribonucleoprotein (U2snRNP). Metazoan gene duplication further specialized shared TFIID/SAGA subunits into SAGA-specific paralogs (TAF5L and TAF6L), but the functional consequences of this divergence are unknown. We report the structure of endogenous human SAGA purified via an affinity ligand from cells that were not disturbed by any genomic engineering tools. Our work reveals the high-resolution structure of SPL and the TAF6L HEAT repeat domain that provides the SPL with a docking surface. We elucidate how SPL and the HEAT repeats are incorporated into SAGA. We identify major structural differences between TAF6L/TAF5L and their canonical paralogs that enable SPL accommodation. SPL engages SAGA through a substantially smaller interface than in U2snRNP, despite sharing a deeply inserted helical motif. The seemingly weaker interaction of SPL with SAGA raises the possibility that SAGA relays this module to the splicing machinery.

Structural insights explain how human SAGA evolved to host a splicing module absent in yeast.

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