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Target Discrimination in Nonsense-Mediated mRNA Decay Requires Upf1 ATPase Activity.

Molecular cell · 2015 · Vol. 59 (3) · pp. 413-25

Abstract

RNA quality-control pathways get rid of faulty RNAs and therefore must be able to discriminate these RNAs from those that are normal. Here we present evidence that the adenosine triphosphatase (ATPase) cycle of the SF1 helicase Upf1 is required for mRNA discrimination during nonsense-mediated decay (NMD). Mutations affecting the Upf1 ATPase cycle disrupt the mRNA selectivity of Upf1, leading to indiscriminate accumulation of NMD complexes on both NMD target and non-target mRNAs. In addition, two modulators of NMD-translation and termination codon-proximal poly(A) binding protein-depend on the ATPase activity of Upf1 to limit Upf1-non-target association. Preferential ATPase-dependent dissociation of Upf1 from non-target mRNAs in vitro suggests that selective release of Upf1 contributes to the ATPase dependence of Upf1 target discrimination. Given the prevalence of helicases in RNA regulation, ATP hydrolysis may be a widely used activity in target RNA discrimination.

Publication Types

["Journal Article", "Research Support, N.I.H., Extramural", "Research Support, Non-U.S. Gov't"]

Keywords

[]

MeSH Terms

["3' Untranslated Regions", "Adenosine Triphosphate", "Catalytic Domain", "HEK293 Cells", "Humans", "In Vitro Techniques", "Molecular Sequence Data", "Mutation", "Nonsense Mediated mRNA Decay", "RNA Helicases", "RNA, Messenger", "Substrate Specificity", "Trans-Activators"]

Funding

R01 HG004659 NHGRI NIH HHS (United States)
T32 GM008666 NIGMS NIH HHS (United States)
U54 HG007005 NHGRI NIH HHS (United States)
R01 NS075449 NINDS NIH HHS (United States)
HG004659 NHGRI NIH HHS (United States)
NS075449 NINDS NIH HHS (United States)
HG007005 NHGRI NIH HHS (United States)
R01 GM084317 NIGMS NIH HHS (United States)
R01 GM099717 NIGMS NIH HHS (United States)

Linked Datasets (3)

GSE69584 GSE via ncbi_elink
GEO

Target discrimination in nonsense-mediated mRNA decay requires Upf1 ATPase activity (CLIP-Seq)

Homo sapiens
12 data files
FileTypeSize
FJM_W1_NoIndex_L001_R1.fastq.gz OTHER 4.6 GB
FJM_W1_NoIndex_L001_R1.fastq.gz OTHER 4.6 GB
FM_UPF_1_NoIndex_L004_R1.fastq.gz OTHER 6.5 GB
FM_UPF_1_NoIndex_L004_R1.fastq.gz OTHER 6.5 GB
FM_UPF_2_NoIndex_L005_R1.fastq.gz OTHER 6.8 GB
FM_UPF_2_NoIndex_L005_R1.fastq.gz OTHER 6.8 GB
FM_UPF_3_NoIndex_L006_R1.fastq.gz OTHER 7.7 GB
FM_UPF_3_NoIndex_L006_R1.fastq.gz OTHER 7.7 GB
FM_UPF_4_NoIndex_L007_R1.fastq.gz OTHER 6.8 GB
SRR2051425 OTHER 6.8 GB
WT_NoIndex_L006_R1.fastq.gz OTHER 6.5 GB
WT_NoIndex_L006_R1.fastq.gz OTHER 6.5 GB
GSE69586 GSE via ncbi_elink
GEO

Target discrimination in nonsense-mediated mRNA decay requires Upf1 ATPase activity

Homo sapiens
GSE69585 GSE via ncbi_elink
GEO

Target discrimination in nonsense-mediated mRNA decay requires Upf1 ATPase activity (RIP-Seq)

Homo sapiens
16 data files
FileTypeSize
D-in_AGCGATAG-TATAGCCT_L001_R1.fastq.gz RIP-Seq 1.7 GB
D-IP_ATTACTCG-ATAGAGGC_L001_R1.fastq.gz RIP-Seq 5.3 GB
D-IP_ATTACTCG-ATAGAGGC_L001_R1.fastq.gz RIP-Seq 5.3 GB
F-in_TAATGCGC-TATAGCCT_L004_R1.fastq.gz RIP-Seq 497.7 MB
F-in_TAATGCGC-TATAGCCT_L004_R1.fastq.gz RIP-Seq 497.7 MB
F-IP_CGGCTATG-TATAGCCT_L007_R1.fastq.gz RIP-Seq 317.5 MB
F-IP_CGGCTATG-TATAGCCT_L007_R1.fastq.gz RIP-Seq 317.5 MB
K-in_TCCGGAGA-ATAGAGGC_L001_R1.fastq.gz RIP-Seq 1.6 GB
K-in_TCCGGAGA-ATAGAGGC_L001_R1.fastq.gz RIP-Seq 1.6 GB
K-IP_CGCTCATT-ATAGAGGC_L001_R1.fastq.gz RIP-Seq 1.0 GB
SRR2051427 RIP-Seq 1.7 GB
SRR2051432 RIP-Seq 1.0 GB
SRR2051433 RIP-Seq 1.1 GB
SRR2051434 RIP-Seq 508.1 MB
W-in_TCCGCGAA-TATAGCCT.fastq.gz RIP-Seq 1.1 GB
W-IP_TCTCGCGC-TATAGCCT_L001_R1.fastq.gz RIP-Seq 508.1 MB

Analysis Pipelines (3)

CLIP-seq geo_data_processing GSE69584
CLIP-seq geo_data_processing GSE69585
CLIP-seq geo_data_processing GSE69586