The C terminus of Pcf11 forms a novel zinc-finger structure that plays an essential role in mRNA 3′-end processing

Yang, Fan; Hsu, Peter L.; Lee, Susan D.; Yang, Wen; Hoskinson, Derick; Xu, Weihao; Moore, Claire; Varani, Gabriele

doi:10.1261/rna.058354.116

Cited by 20 publications

(20 citation statements)

References 42 publications

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“…This would be expected to result in a longer disordered loop, but otherwise maintain all other characteristics of this zinc-binding motif. The putative C-terminal zinc finger domain of VPS39 is significantly shorter than those of VPS18 and VPS41, containing only four Zn 2+ ligands, and the closest homologue of known structure is the zinc finger domain of yeast Pcf11 (42).…”

Section: Resultsmentioning

confidence: 97%

VPS18 recruits VPS41 to the human HOPS complex via a RING-RING interaction

Hunter

Scourfield

Emmott

et al. 2017

Preprint

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The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/169185 doi: bioRxiv preprint first posted online 2 ABSTRACT Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET and HOPS, to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date there have been far fewer studies of metazoan CORVET and HOPS. Both of these complexes comprise six subunits: a common four-subunit core and two unique subunits. Once assembled, these complexes function to recognise specific endosomal membrane markers and facilitate SNARE-mediated membrane fusion.CORVET promotes the homotypic fusion of early endosomes, while HOPS promotes the fusion of lysosomes to late endosomes and autophagosomes. Many of the subunits of both CORVET and HOPS contain putative C-terminal zinc-finger domains. Here, the contribution of these domains to the assembly of the human CORVET and HOPS complexes has been examined. Using biochemical techniques, we demonstrate that the zinc-containing RING domains of human VPS18 and VPS41 interact directly to form a stable heterodimer. In cells, these RING domains are able to integrate into endogenous HOPS, showing that the VPS18 RING domain is required to recruit VPS41 to the core complex subunits. Importantly, this mechanism is not conserved throughout eukaryotes, as yeast Vps41 does not contain a C-terminal zinc-finger motif. The subunit analogous to VPS41 in human CORVET is VPS8, in which the RING domain has an additional C-terminal segment that is predicted to be disordered. Both the RING and disordered C-terminal domains are required for integration of VPS8 into endogenous CORVET complexes, suggesting that HOPS and CORVET recruit VPS41 and VPS8 via distinct molecular interactions.

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Section: Resultsmentioning

confidence: 97%

VPS18 recruits VPS41 to the human HOPS complex via a RING-RING interaction

Hunter

Scourfield

Emmott

et al. 2017

Preprint

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show abstract

“…This would be expected to result in a longer disordered loop, but otherwise maintain all other characteristics of this zinc-binding motif. The putative C-terminal zinc-finger domain of VPS39 is significantly shorter than that of VPS18 and VPS41, containing only four Zn 2+ ligands, and the closest homologue of known structure is the zinc-finger domain of yeast Pcf11 [ 42 ].…”

Section: Resultsmentioning

confidence: 99%

VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction

Hunter

Scourfield

Emmott

et al. 2017

Biochemical Journal

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Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET (class C core vacuole/endosome tethering) and HOPS (homotypic fusion and vacuole protein sorting), to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date there have been far fewer studies of metazoan CORVET and HOPS. Both of these complexes comprise six subunits: a common four-subunit core and two unique subunits. Once assembled, these complexes function to recognise specific endosomal membrane markers and facilitate SNARE-mediated membrane fusion. CORVET promotes the homotypic fusion of early endosomes, while HOPS promotes the fusion of lysosomes to late endosomes and autophagosomes. Many of the subunits of both CORVET and HOPS contain putative C-terminal zinc-finger domains. Here, the contribution of these domains to the assembly of the human CORVET and HOPS complexes has been examined. Using biochemical techniques, we demonstrate that the zinc-containing RING (really interesting new gene) domains of human VPS18 and VPS41 interact directly to form a stable heterodimer. In cells, these RING domains are able to integrate into endogenous HOPS, showing that the VPS18 RING domain is required to recruit VPS41 to the core complex subunits. Importantly, this mechanism is not conserved throughout eukaryotes, as yeast Vps41 does not contain a C-terminal zinc-finger motif. The subunit analogous to VPS41 in human CORVET is VPS8, in which the RING domain has an additional C-terminal segment that is predicted to be disordered. Both the RING and disordered C-terminal domains are required for integration of VPS8 into endogenous CORVET complexes, suggesting that HOPS and CORVET recruit VPS41 and VPS8 via distinct molecular interactions.

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“…Numerous dimethylarginine residues in the repeats were also detected in the MS analysis of our baculovirus-produced hPcf11 (data not shown). The C-terminus of Pcf11 contains two zinc fingers (Barillà et al 2001;Sadowski et al 2003;Guéguéniat et al 2017;Yang et al 2017), which straddle the Clp1 interaction site (Noble et al 2007). The region of yPcf11 responsible for the Rna14/Rna15 interaction is on the N-terminal side of the first zinc finger (Amrani et al 1997) and has been narrowed down to amino acids 331-417 (Lionel Minvielle-Sebastia, pers.…”

Section: Domain Organization Of Cf IImentioning

confidence: 99%

Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors

Schäfer

Tüting

Schönemann

et al. 2018

RNA

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Cleavage factor II (CF II) is a poorly characterized component of the multiprotein complex catalyzing 3' cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reactions, whereas hClp1 by itself is not. Pcf11 moderately stimulates the RNA 5' kinase activity of hClp1; the kinase activity is dispensable for RNA cleavage. CF II binds RNA with nanomolar affinity. Binding is mediated mostly by the two zinc fingers in the C-terminal region of hPcf11. RNA is bound without pronounced sequence-specificity, but extended G-rich sequences appear to be preferred. We discuss the possibility that CF II contributes to the recognition of cleavage/polyadenylation substrates through interaction with G-rich far-downstream sequence elements.

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The C terminus of Pcf11 forms a novel zinc-finger structure that plays an essential role in mRNA 3′-end processing

Cited by 20 publications

References 42 publications

VPS18 recruits VPS41 to the human HOPS complex via a RING-RING interaction

VPS18 recruits VPS41 to the human HOPS complex via a RING-RING interaction

VPS18 recruits VPS41 to the human HOPS complex via a RING–RING interaction

Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors

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