Non-gradient and genotype-dependent patterns of RSV gene expression


Autoři: Felipe-Andrés Piedra aff001;  Xueting Qiu aff002;  Michael N. Teng aff003;  Vasanthi Avadhanula aff001;  Annette A. Machado aff001;  Do-Kyun Kim aff004;  James Hixson aff004;  Justin Bahl aff002;  Pedro A. Piedra aff001
Působiště autorů: Department of Molecular Virology & Microbiology, Baylor College of Medicine, Houston, TX, United States of America aff001;  Center for the Ecology of Infectious Diseases, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA, United States of America aff002;  Division of Allergy and Immunology, Department of Internal Medicine, University of South Florida Morsani College of Medicine, Tampa, FL, United States of America aff003;  Human Genetics Center, School of Public Health, University of Texas Health Science Center, Houston, TX, United States of America aff004;  Program in Emerging Infectious Diseases, Duke-National University of Singapore Graduate Medical School, Singapore aff005;  Department of Pediatrics, Baylor College of Medicine, Houston, TX, United States of America aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pone.0227558

Souhrn

Respiratory syncytial virus (RSV) is a nonsegmented negative-strand RNA virus (NSV) and a leading cause of severe lower respiratory tract illness in infants and the elderly. Transcription of the ten RSV genes proceeds sequentially from the 3’ promoter and requires conserved gene start (GS) and gene end (GE) signals. Previous studies using the prototypical GA1 genotype Long and A2 strains have indicated a gradient of gene transcription extending across the genome, with the highest level of mRNA coming from the most promoter-proximal gene, the first nonstructural (NS1) gene, and mRNA levels from subsequent genes dropping until reaching a minimum at the most promoter-distal gene, the polymerase (L) gene. However, recent reports show non-gradient levels of mRNA, with higher than expected levels from the attachment (G) gene. It is unknown to what extent different transcript stabilities might shape measured mRNA levels. It is also unclear whether patterns of RSV gene expression vary, or show strain- or genotype-dependence. To address this, mRNA abundances from five RSV genes were measured by quantitative real-time PCR (qPCR) in three cell lines and in cotton rats infected with RSV isolates belonging to four genotypes (GA1, ON, GB1, BA). Relative mRNA levels reached steady-state between four and 24 hours post-infection. Steady-state patterns were non-gradient and genotype-specific, where mRNA levels from the G gene exceeded those from the more promoter-proximal nucleocapsid (N) gene across isolates. Transcript stabilities could not account for the non-gradient patterns observed, indicating that relative mRNA levels more strongly reflect transcription than decay. Our results indicate that gene expression from a small but diverse set of RSV genotypes is non-gradient and genotype-dependent. We propose novel models of RSV transcription that can account for non-gradient transcription.

Klíčová slova:

DNA transcription – Gene expression – Genomic signal processing – Messenger RNA – Oligonucleotides – Polymerases – Transcriptional control – Viral gene expression


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