Dataset Information

ARIH2 is a Vif-dependent regulator of CUL5-mediated APOBEC3G degradation in HIV infection: SRM-CBFB

ABSTRACT: SRM assays were generated for selected interactors of CBFB. SRM assay generation was performed using Skyline. For all targeted proteins, proteotypic peptides and optimal transitions for identification and quantification were selected based on the Skyline spectral library generated from the shotgun MS experiments (MSV000084855). For each protein, 2-5 peptides were selected based on intensity, peptide length as well as chromatographic performance. For each peptide the 4 best SRM transitions were selected based on intensity and peak shape. Digested peptide mixtures were analyzed by LC-SRM on a Thermo Scientific TSQ Quantiva MS system equipped with a Proxeon. Easy nLC 1200 ultra high-pressure liquid chromatography and autosampler system. Samples were injected onto a C18 column (25 cm x 75 mm I.D. packed with ReproSil Pur C18 AQ 1.9 mm particles) in 0.1% formic acid and then separated with an 80 min gradient from 5% to 40% Buffer B (90% ACN/10% water/0.1% formic acid) at a flow rate of 300 nL/min. SRM acquisition was per- formed operating Q1 and Q3 at 0.7 unit mass resolution. For each peptide the best 4 transitions were monitored in a scheduled fashion with a retention time window of 4 min and a cycle time fixed to 2 seconds. Argon was used as the collision gas at a nominal pressure of 1.5 mTorr. Collision energies were calculated by, CE = 0.0348 * (m/z) + 0.4551 and CE = 0.0271 * (m/z) + 1.5910 (CE, collision energy and m/z, mass to charge ratio) for doubly and triply charged precursor ions, respectively. RF lens voltages were calculated by, RF = 0.1088 * (m/z) + 21.029 and RF = 0.1157 * (m/z) + 0.1157 (RF, RF lens voltage and m/z, mass to charge ratio) for doubly and triply charged precursor ions, respectively. The resulting data was analyzed with Skyline for identification and quantification of peptides. MSstats was used for statistical analysis.

INSTRUMENT(S): TSQ Quantiva

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Human Immunodeficiency Virus 1 (ncbitaxon:11676)

SUBMITTER: Nevan J. Krogan

PROVIDER: MSV000084964 | MassIVE | Mon Feb 17 17:31:00 GMT 2020

SECONDARY ACCESSION(S): PXD017566

REPOSITORIES: MassIVE

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ARIH2 Is a Vif-Dependent Regulator of CUL5-Mediated APOBEC3G Degradation in HIV Infection.

Hüttenhain Ruth R Xu Jiewei J Burton Lily A LA Gordon David E DE Hultquist Judd F JF Johnson Jeffrey R JR Satkamp Laura L Hiatt Joseph J Rhee David Y DY Baek Kheewoong K Crosby David C DC Frankel Alan D AD Marson Alexander A Harper J Wade JW Alpi Arno F AF Schulman Brenda A BA Gross John D JD Krogan Nevan J NJ

Cell host & microbe 20190625 1

The Cullin-RING E3 ligase (CRL) family is commonly hijacked by pathogens to redirect the host ubiquitin proteasome machinery to specific targets. During HIV infection, CRL5 is hijacked by HIV Vif to target viral restriction factors of the APOBEC3 family for ubiquitination and degradation. Here, using a quantitative proteomics approach, we identify the E3 ligase ARIH2 as a regulator of CRL5-mediated APOBEC3 degradation. The CUL5Vif/CBFß complex recruits ARIH2 where it acts to transfer ...[more]

PMID: 31253590

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