Respiratory syncytial virus M2-1 protein associates non-specifically with viral messenger RNA and with specific cellular messenger RNA transcripts.

Abstract

Respiratory syncytial virus (RSV) is a major cause of respiratory disease in infants and the elderly. RSV is a non-segmented negative strand RNA virus. The viral M2-1 protein plays a key role in viral transcription, serving as an elongation factor to enable synthesis of full-length mRNAs. M2-1 contains an unusual CCCH zinc-finger motif that is conserved in the related human metapneumovirus M2-1 protein and filovirus VP30 proteins. Previous biochemical studies have suggested that RSV M2-1 might bind to specific virus RNA sequences, such as the transcription gene end signals or poly A tails, but there was no clear consensus on what RSV sequences it binds. To determine if M2-1 binds to specific RSV RNA sequences during infection, we mapped points of M2-1:RNA interactions in RSV-infected cells at 8 and 18 hours post infection using crosslinking immunoprecipitation with RNA sequencing (CLIP-Seq). This analysis revealed that M2-1 interacts specifically with positive sense RSV RNA, but not negative sense genome RNA. It also showed that M2-1 makes contacts along the length of each viral mRNA, indicating that M2-1 functions as a component of the transcriptase complex, transiently associating with nascent mRNA being extruded from the polymerase. In addition, we found that M2-1 binds specific cellular mRNAs. In contrast to the situation with RSV mRNA, M2-1 binds discrete sites within cellular mRNAs, with a preference for A/U rich sequences. These results suggest that in addition to its previously described role in transcription elongation, M2-1 might have an additional role involving cellular RNA interactions.

• Authors: Molly R Braun, Sarah L Noton, Emmeline L Blanchard, Afzaal Shareef, Philip J Santangelo, W Evan Johnson, Rachel Fearns
• Journal: PLoS pathogens (PLoS Pathog) Vol. 17 Issue 5 Pg. e1009589 (05 2021) ISSN: 1553-7374 [Electronic] United States
• PMID: 34003848 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• RNA, Messenger
• RNA-Binding Proteins
• Transcriptional Elongation Factors
• Viral Proteins
• human respiratory syncytial virus M2-1 protein
• DNA-Directed RNA Polymerases

Topics

• DNA-Directed RNA Polymerases (genetics, metabolism)
• Humans
• RNA, Messenger (genetics, metabolism)
• RNA-Binding Proteins
• Respiratory Syncytial Virus Infections (virology)
• Respiratory Syncytial Virus, Human (genetics)
• Transcriptional Elongation Factors (genetics, metabolism)
• Viral Proteins (genetics, metabolism)
• Virus Replication