IndraLab

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18 | 9 16

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"In the nucleus, PRPF8 is forming a complex with EFTUD2 and SNRNP200 (also known as BRR2) as part of the spliceosome."

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"The conservative missense variant p.PRPF8-Arg2310Lys that is not altering the global charge of the C-terminal tail, and variants located at the basis of the C-terminal tail show milder clinical phenotypes, in accordance with functional data on PRPF8 and SNRNP200 interactions in yeast."

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"For example, pathogenic variants in PRPF8 and SNRNP200 were clustered in their interaction regions, where defects disrupted PRPF8SNRNP200 interactions. xref – xref Previously, situations similar to that of TOPORS have been reported in the RP1 gene, in which causative heterozygous truncating variants are only located in a specific region in the middle of the gene. xref – xref Regarding TOPORS , products resulting from pathogenic truncating variants located in the last exon and downstream of the SR/RS domain may escape nonsense-mediated mRNA decay (NMD) xref and reduce the activity of the wild-type allele via the dominant-negative effect."

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"Furthermore, TSSC4 directly interacts with the SNRNP200 (U5 snRNP 200 kDa helicase) and the PRPF8 spliceosomal scaffold."

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"In addition, a small but reproducible reduction of SNRNP200 and PRPF8 binding was also observed in the case of the Δ201–250 deletion mutant."

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"These Prp8-RP mutants directly affect U5 snRNP and tri-snRNP maturation (Boon et al. 2007), Brr2 helicase, and ATPase activities (Pena et al. 2007; Maeder et al. 2009; Mozaffari-Jovin et al. 2013), and yeast-two-hybrid interactions between Brr2, Prp8, and Snu114 (Table 1; Pena et al. 2007)."

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"To address this question, we used enhanced (e)CLIP xref , xref to monitor the association of snRNA variants with spliceosome components SNRNP200 (BRR2) and PRPF8 (PRP8) in the absence or presence of NEXT complex depletion ( xref and xref – xref )."

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"Since neither PRPF8 nor SNRNP200 interact directly with U1 snRNA, we used formaldehyde- rather than UV-crosslinking to ensure capture of the entire spliceosome complex."

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"We found that for a majority of observed U1 snRNA variants, NEXT complex depletion resulted in a 10- to 50-fold increase in their association with SNRNP200 and PRPF8 as compared with control depletion conditions ( xref and xref )."

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"It suggests that the C-terminal tail of PRPF8, which is inserted into and blocks the RNA tunnel in SNRNP200, does not considerably contribute to the interaction between PRPF8 and SNRNP200 and overall stability of the U5 snRNP ( xref )."

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"The level of U2 snRNA variants associated with SNRNP200 and PRPF8 also increased upon NEXT depletion ( xref – xref and xref – xref ), mostly due to one U2 variant, RNU2–63P ( xref )."

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"BRR2 interacts extensively with the U5-specific protein PRPF8 [ xref ]."

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"SNRNP200 tightly interacts with the splicing factor PRPF8, and these two proteins have been shown to assemble in the cytoplasm and be transported to the nucleus ( xref )."

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"All the known variants cluster in the C-terminal Jab1/MPN domain of the PRPF8 protein which interacts with SNRNP200 ( xref ; xref ; xref ; xref ; xref ; xref )."

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"Moreover, they also report that a loss of PHF6 results in increased genomic instability at rDNA genes and a cell cycle delay at G2/M. While this was the first indication of a nucleolar function for PHF6, it has also been reported to interact with PRPF8 and SNRNP200, both constituents of the U4/U6-U5 tri-snRNP pre-mRNA splicing ribonucleoprotein complex [ xref ]."

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"CryoEM structure of a BRR2-PRPF8 Jab1 -C9ORF78 complex."

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"Numerous structural studies show that SNRNP200 and PRPF8 form a complex that stimulates SNRNP200 unwinding activity (Malinová et al. 2017; Preussner et al. 2022; Bergfort, Hilal, et al. 2022; Bergfort, Preußner, et al. 2022)."

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"2 ▸ g and 2 ▸ h) and failed to sequester PRPF8 from a pre-formed SNRNP200PRPF8 complex (Fig. 3 ▸ a and 3 ▸ b)."

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"However, neither of the two Prpf8 aberrant mutations studied in this work affect the Prpf8 interaction with Prpf6 and Snrnp200 and snRNP maturations (Malinova et al, 2017, and Fig S1C)."

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"It suggests that the C-terminal tail of PRPF8, which is inserted into and blocks the RNA tunnel in SNRNP200, does not considerably contribute to the interaction between PRPF8 and SNRNP200 and overall stability of the U5 snRNP (Mozaffari-Jovin et al, 2013)."

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"AAR2 alone or in complex with PRPF8 did not bind stably to SNRNP200 or to a SNRNP200PRPF8 complex (Figs."

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"We hypothesize that the studied RP mutations alter the interaction of PRPF8 and SNRNP200 with RNA."

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"2 xref g and 2 xref h ) and failed to sequester PRPF8 JM from a pre-formed SNRNP200 395–2136 –PRPF8 JM complex (Fig. 3 xref a and 3 xref b )."

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"We hypothesize that the studied RP mutations alter the interaction of PRPF8 and SNRNP200 with RNA."

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"AAR2 alone or in complex with PRPF8 RH did not bind stably to SNRNP200 395–2136 or to a SNRNP200 395–2136 –PRPF8 JM complex (Figs."

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