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The effect of TuMV on translation initiation in Arabidopsis

ABSTRACT: Arabidopsis ecotype Col-0 expressing RPL18 fused to His & Flag epitopes (RPL18-HF) under 35S control was used to discern the effects of Turnip mosaic virus (TuMV) infection on host translation initiation. Plants were grown in LC-1 soil in 12 hour days and infected by TuMV-GFP or mock-inoculated just prior to sending up bolts. Samples were taken from rosette leaves 10 days after inoculation. Only tissue fluorescing GFP was selected from the virus-infected samples. Similar tissue was sampled from mock-infected leaves. FLAG antibody was used to isolate RPL18-HF. The RNA co-immunoprecipitated with RPL18-HF is fully translation-initiated. This translation-initiated RNA, also referred to as polysomal RNA, was isolated and compared to total RNA under both mock and TuMV-infected states to find TuMV-induced changes in host translation initiation. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Jackson Moeller. The equivalent experiment is AT42 at PLEXdb.] pathogen infection: TuMV inoculated - RNA fraction: polysomal RNA(3-replications); pathogen infection: TuMV inoculated - RNA fraction: total RNA(3-replications); pathogen infection: mock inoculated - RNA fraction: polysomal RNA(3-replications); pathogen infection: mock inoculated - RNA fraction: total RNA(3-replications)

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Sudhansu PLEXdb Curator

PROVIDER: E-GEOD-20278 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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The effect of TuMV on translation initiation in Arabidopsis

Project description:Arabidopsis ecotype Col-0 expressing RPL18 fused to His & Flag epitopes (RPL18-HF) under 35S control was used to discern the effects of Turnip mosaic virus (TuMV) infection on host translation initiation. Plants were grown in LC-1 soil in 12 hour days and infected by TuMV-GFP or mock-inoculated just prior to sending up bolts. Samples were taken from rosette leaves 10 days after inoculation. Only tissue fluorescing GFP was selected from the virus-infected samples. Similar tissue was sampled from mock-infected leaves. FLAG antibody was used to isolate RPL18-HF. The RNA co-immunoprecipitated with RPL18-HF is fully translation-initiated. This translation-initiated RNA, also referred to as polysomal RNA, was isolated and compared to total RNA under both mock and TuMV-infected states to find TuMV-induced changes in host translation initiation. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Jackson Moeller. The equivalent experiment is AT42 at PLEXdb.]

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Project description:The goal of this experiment was to examine the effects of Blumeria gramins f. sp. hordei (Bgh) on the host process of translation initiation in both incompatible and compatible interactions with Hordeum vulgare. This was done by using Bgh isolate 5874 (AVRa6) on the resistant host CI 16151 (Mla6) and fast-neutron derived, loss-of-function mutant m9472 (mla6). Plants were grown 7 days in the greenhouse and then inoculated or non-inoculated and transferred to an 18°C growth chamber with an 8 hr dark, 16 hr light photoperiod. At 32 hours after infection the tissue was harvested and immediately frozen in liquid nitrogen. From the same tissue, total RNA was isolated using a Trizol method, while polysomal RNA was isolated by sucrose density centrifugation. Three biological replications were conducted. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Jackson Moeller. The equivalent experiment is BB72 at PLEXdb.] genotype:CI 16151 (Mla6)-pathogen infection: Bgh isolate 5874 (AVRa6)-RNA fraction:polysomal RNA(3-reps); genotype:CI 16151 (Mla6)-pathogen infection: Bgh isolate 5874 (AVRa6)-RNA fraction:total RNA(3-reps); genotype:CI 16151 (Mla6)-pathogen infection:non-inoculated-RNA fraction:polysomal RNA(3-reps); genotype:CI 16151 (Mla6)-pathogen infection:non-inoculated-RNA fraction:total RNA(3-reps); genotype:CI 16151 m9472 (mla6)-pathogen infection: Bgh isolate 5874 (AVRa6)-RNA fraction:polysomal RNA(3-reps); genotype:CI 16151 m9472 (mla6)-pathogen infection: Bgh isolate 5874 (AVRa6)-RNA fraction:total RNA(3-reps); genotype:CI 16151 m9472 (mla6)-pathogen infection:non-inoculated-RNA fraction:polysomal RNA(3-reps); genotype:CI 16151 m9472 (mla6)-pathogen infection:non-inoculated-RNA fraction:total RNA(3-reps)

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