Co-regulatory activity of hnRNP K and NS1-BP in influenza and human mRNA splicing

Matthew G Thompson; Raquel Muñoz-Moreno; Prasanna Bhat; Renat Roytenberg; John Lindberg; Matthew R Gazzara; Michael J Mallory; Ke Zhang; Adolfo García-Sastre; Beatriz M A Fontoura; Kristen W Lynch

doi:10.1038/s41467-018-04779-4

Co-regulatory activity of hnRNP K and NS1-BP in influenza and human mRNA splicing

Nat Commun. 2018 Jun 19;9(1):2407. doi: 10.1038/s41467-018-04779-4.

Authors

Affiliations

¹ Departments of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, 422 Curie Blvd, Philadelphia, PA, 19104, USA.
² Department of Microbiology, Icahn School of Medicine at Mount Sinai, 1468 Madison Ave., Box 1124, New York, NY, 10029, USA.
³ Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, 1468 Madison Ave., Box 1124, New York, NY, 10029, USA.
⁴ Department of Cell Biology, UT Southwestern Medical Center, 6000 Harry Hines Blvd., Dallas, TX, 75390, USA.
⁵ Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, 1468 Madison Ave., Box 1124, New York, NY, 10029, USA.
⁶ Departments of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, 422 Curie Blvd, Philadelphia, PA, 19104, USA. klync@pennmedicine.upenn.edu.

Abstract

Three of the eight RNA segments encoded by the influenza A virus (IAV) undergo alternative splicing to generate distinct proteins. Previously, we found that host proteins hnRNP K and NS1-BP regulate IAV M segment splicing, but the mechanistic details were unknown. Here we show NS1-BP and hnRNP K bind M mRNA downstream of the M2 5' splice site (5'ss). NS1-BP binds most proximal to the 5'ss, partially overlapping the U1 snRNP binding site, while hnRNP K binds further downstream and promotes U1 snRNP recruitment. Mutation of either or both the hnRNP K and NS1-BP-binding sites results in M segment mis-splicing and attenuated IAV replication. Additionally, we show that hnRNP K and NS1-BP regulate host splicing events and that viral infection causes mis-splicing of some of these transcripts. Therefore, our proposed mechanism of hnRNP K/NS1-BP mediated IAV M splicing provides potential targets of antiviral intervention and reveals novel host functions for these proteins.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Heterogeneous-Nuclear Ribonucleoprotein K / genetics*
Heterogeneous-Nuclear Ribonucleoprotein K / metabolism
Humans
Influenza A virus / genetics
Influenza, Human / genetics*
Influenza, Human / metabolism
Influenza, Human / virology
Mutation
Nuclear Proteins / genetics*
Nuclear Proteins / metabolism
RNA Precursors / genetics*
RNA Precursors / metabolism
RNA Splicing*
RNA, Messenger / genetics*
RNA, Messenger / metabolism
RNA, Viral / genetics
RNA, Viral / metabolism
RNA-Binding Proteins
Ribonucleoprotein, U1 Small Nuclear / genetics
Ribonucleoprotein, U1 Small Nuclear / metabolism
Transcription Factors / genetics*
Transcription Factors / metabolism
Virus Replication / genetics

Substances

Heterogeneous-Nuclear Ribonucleoprotein K
IVNS1ABP protein, human
Nuclear Proteins
RNA Precursors
RNA, Messenger
RNA, Viral
RNA-Binding Proteins
Ribonucleoprotein, U1 Small Nuclear
Transcription Factors

Abstract

Publication types

MeSH terms

Substances

Grants and funding