التفاصيل البيبلوغرافية
| العنوان: |
Characterization of endogenous MECR transcripts and cMECR:polymerase interactions. |
| المؤلفون: |
Steven F. Baker (847771), Helene Meistermann (14290500), Manuel Tzouros (2373868), Aaron Baker (2162533), Sabrina Golling (11831093), Juliane Siebourg Polster (14290503), Mitchell P. Ledwith (14290506), Anthony Gitter (142451), Angelique Augustin (11831096), Hassan Javanbakht (2403709), Andrew Mehle (655696) |
| سنة النشر: |
2022 |
| مصطلحات موضوعية: |
Biochemistry, Microbiology, Cell Biology, Molecular Biology, Biotechnology, Evolutionary Biology, Cancer, Infectious Diseases, Virology, Biological Sciences not elsewhere classified, yeast homolog etr1, used immunocompetitive capture, frequently switch hosts, cryptic cytoplasmic isoform, alternative splicing produces, alternative splicing liberates, viral ribonucleoprotein complexes, style viral polymerases, cryptic antiviral protein, avoiding host defense, mitochondrial enoyl coa, identify cellular proteins, host cell machinery, specific antiviral activity, causing localized outbreaks, suppresses viral replication, antiviral activity, viral polymerase, host range, cellular partners |
| الوصف: |
(A-B) RT-PCR analysis of poly-adenylated MECR transcripts in mock or WSN-infected A549 cells (MOI 1, 6 h). ( A ) Genomic representation of MECR and regions amplified by specific primer sets. Untranslated regions (UTRs) in dark gray boxes, exons in light gray boxes, and introns in lines. Diagram segments are not to scale; base pair (bp) lengths are indicated. ( B ) Amplification of transcripts containing exon 1 (red), a region spanning exons 1 and 2 (green), and the unique cMECR UTR through exon 3 (yellow). The primer pair MECR ex1 FOR/MECR ex2 REV amplifies MECR (205 bp, empty circle) and cMECR (332 bp, filled circle). cMECR is predicted to be variable lengths around 332 bp due to the use of multiple splice donor sites paired with the cMECR UTR splice acceptor site. Asterisk; nonspecific band. Size shown in bp derived from a DNA ladder. ( C ) Ribosomes initiate on the short splice isoform transcript in the UTR and at the cMECR start site. Cells were treated with LTM to identify initiation sites during ribosome profiling (data from [ 51 ]). RPFs and total RNA (RNA-Seq) were mapped to the MECR locus, showing initiation events at the predicted cMECR start codon and other sites in the short splice isoform encoding cMECR. Zoom on exon 2 shows coding potential of the short isoform with stop codons in red, start codons in green, and the cMECR initiation site with an arrow. (D) PB2-FLAG and V5-tagged MECR or cMECR were expressed in 293T cells with or without the other polymerase (PB1/PA) or vRNP (vNA/NP) components. Cells were lysed and immunoprecipitated with anti-V5 antibody or IgG controls. Proteins were detected by western blot. ( E ) Subcellular localization of mCh fusion constructs. Plasmids encoding codon-optimized MECR (M77L) or cMECR fused to mCh with a 5GS linker were transfected into 293T cells. Red fluorescence images were captured 24 h posttransfection. Scale bars; 20 μm. ( F ) cMECR-AirID biotinylates polymerase. Cells coexpressing PB2-FLAG-tagged polymerase and free AirID or the cMECR-AirID fusion were ... |
| نوع الوثيقة: |
still image |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/figure/Characterization_of_endogenous_i_MECR_i_transcripts_and_cMECR_polymerase_interactions_/21766041 |
| DOI: |
10.1371/journal.pbio.3001934.s005 |
| الاتاحة: |
https://doi.org/10.1371/journal.pbio.3001934.s005 |
| Rights: |
CC BY 4.0 |
| رقم الانضمام: |
edsbas.A33646E7 |
| قاعدة البيانات: |
BASE |