8R8R

Cryo-EM structure of the human mPSF with PAPOA C-terminus peptide (PAPOAc)

8R8R の概要

• エントリーDOI: 10.2210/pdb8r8r/pdb
• EMDBエントリー: 19008
• 分子名称: Cleavage and polyadenylation specificity factor subunit 1, pre-mRNA 3' end processing protein WDR33, Cleavage and polyadenylation specificity factor subunit 4, ... (6 entities in total)
• 機能のキーワード: 3-end processing, polyadenylation, papoa, pre-mrna, cleavage and polyadenylation, rna binding protein
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 245363.66

構造登録者

Todesca, S.,Sandmeir, F.,Keidel, A.,Conti, E. (登録日: 2023-11-29, 公開日: 2024-04-10, 最終更新日: 2024-06-26)

主引用文献

Todesca, S.,Sandmeir, F.,Keidel, A.,Conti, E.
Molecular basis of human poly(A) polymerase recruitment by mPSF.
Rna, 30:795-806, 2024

PubMed Abstract: 3' end processing of most eukaryotic precursor-mRNAs (pre-mRNAs) is a crucial cotranscriptional process that generally involves the cleavage and polyadenylation of the precursor transcripts. Within the human 3' end processing machinery, the four-subunit mammalian polyadenylation specificity factor (mPSF) recognizes the polyadenylation signal (PAS) in the pre-mRNA and recruits the poly(A) polymerase α (PAPOA) to it. To shed light on the molecular mechanisms of PAPOA recruitment to mPSF, we used a combination of cryogenic-electron microscopy (cryo-EM) single-particle analysis, computational structure prediction, and in vitro biochemistry to reveal an intricate interaction network. A short linear motif in the mPSF subunit FIP1 interacts with the structured core of human PAPOA, with a binding mode that is evolutionarily conserved from yeast to human. In higher eukaryotes, however, PAPOA contains a conserved C-terminal motif that can interact intramolecularly with the same residues of the PAPOA structured core used to bind FIP1. Interestingly, using biochemical assay and cryo-EM structural analysis, we found that the PAPOA C-terminal motif can also directly interact with mPSF at the subunit CPSF160. These results show that PAPOA recruitment to mPSF is mediated by two distinct intermolecular connections and further suggest the presence of mutually exclusive interactions in the regulation of 3' end processing.

PubMed: 38538052
DOI: 10.1261/rna.079915.123

実験手法

ELECTRON MICROSCOPY (2.79 Å)