التفاصيل البيبلوغرافية
| العنوان: |
pUL21 recruits PP1 via a conserved motif in the linker region to accelerate CERT dephosphorylation. |
| المؤلفون: |
Tomasz H. Benedyk (11278840), Julia Muenzner (1894084), Viv Connor (3175485), Yue Han (560788), Katherine Brown (3554930), Kaveesha J. Wijesinghe (11278843), Yunhui Zhuang (11242950), Susanna Colaco (41652), Guido A. Stoll (11278846), Owen S. Tutt (11278849), Stanislava Svobodova (11278852), Dmitri I. Svergun (25365), Neil A. Bryant (11278855), Janet E. Deane (3287961), Andrew E. Firth (10325575), Cy M. Jeffries (9173851), Colin M. Crump (7129754), Stephen C. Graham (9904198) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biochemistry, Microbiology, Cell Biology, Biotechnology, Evolutionary Biology, Ecology, Science Policy, Mental Health, Infectious Diseases, Plant Biology, Virology, Environmental Sciences not elsewhere classified, Biological Sciences not elsewhere classified, pUL 21 antagonises, pUS 3 activity, herpes simplex virus replication, herpes simplex virus, virus replication, pUL 21, spread, HSV, protein phosphatase 1, PP 1 recruitment, virus-directed phosphatase activity, pUL 21 PP 1-binding |
| الوصف: |
( A ). Conservation of pUL21 across Alphaverpesvirinae . The following sequences were aligned using ClustalW and conservation calculated using Jalview (Abbreviation and Uniprot ID are shown in parentheses): HSV-1 (HSV1, P10205), HSV-2 (HSV2, G9I242), cercopithecine herpesvirus 2 (CHV2, Q5Y0T2), saimiriine herpesvirus 1 (SHV1, E2IUE9), bovine alphaherpesvirus 1 (BHV1, Q65563), equine herpesvirus 1 (EHV1, P28972), pseudorabies virus (PRV, Q04532), anatid herpesvirus 1 (AHV1, A4GRJ2), varicella-zoster virus (VZV, Q6QCT9), turkey herpesvirus (MHV1, Q9DPR5). Alignment across the linker region (residues 217–280 of HSV-1 pUL21) is shown with conserved residues highlighted. ( B ) HEK293T cells were transfected with plasmids expressing GFP, wild-type (WT) pUL21-GFP or pUL21-GFP with amino acid substitutions in the conserved motif. At 24 hours post-transfection the cells were lysed, subjected to immunoprecipitation using a GFP affinity resin, and captured proteins were subjected to SDS-PAGE and immunoblotting using the listed antibodies. Ponceau S (Pon S) staining of the nitrocellulose membrane before blocking is shown, confirming efficient capture of GFP-tagged proteins. ( C ) Plasmids expressing wild-type or mutant pUL21-GFP, or GFP alone, were transfected into HEK293T cells. At 24 hours post-transfection cells were infected with ΔpUL21 HSV-1 (MOI = 5). Cells were lysed 16 hours post-infection and subjected to immunoprecipitation, SDS-PAGE and immunoblotting as in ( B ). ( D ) Differential scanning fluorimetry of WT (purple) and FV242AA substituted (blue) pUL21-H 6 . Representative curves are shown. Melting temperatures ( T m ) is mean ± standard deviation (n = 3). Inset shows Coomassie-stained SDS-PAGE of the purified protein samples. ( E ) In vitro dephosphorylation assays using all-purified reagents. 0.5 μM CERT was incubated with varying concentrations of GST-PP1 (two-fold serial dilution from 100–3.1 nM) in the absence or presence of 2 μM pUL21-H 6 (WT or FV242AA) for 30 min at 30°C. Proteins were resolved using ... |
| نوع الوثيقة: |
still image |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/figure/pUL21_recruits_PP1_via_a_conserved_motif_in_the_linker_region_to_accelerate_CERT_dephosphorylation_/15174907 |
| DOI: |
10.1371/journal.ppat.1009824.g003 |
| الاتاحة: |
https://doi.org/10.1371/journal.ppat.1009824.g003 |
| Rights: |
CC BY 4.0 |
| رقم الانضمام: |
edsbas.99839B2F |
| قاعدة البيانات: |
BASE |