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Principal Investigator: MELANIE H. COBB
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2024
Award: $6,302
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY (PARENT GRANT)
The goals of this proposal are to uncover how pre-mRNA splicing and nuclear speckles are controlled by
the kinase TAO2, and to determine why TAO2 is required for successful Influenza virus (IAV) replication.
Recent work has identified the understudied protein kinase TAO2 as a host factor essential for splicing and
speckle localization of the IAV M RNA. A pool of TAO2 localizes to nuclear speckles and its loss, by
chemical inhibition or protein depletion, alters nuclear speckle composition, splicing and export of IAV M
RNA, and therefore impairs IAV replication. Inhibition of TAO2 also disrupts splicing of a subset of host
mRNAs, without altering bulk host mRNA. These data uncover a new cellular activity for TAO2 and identify
inhibition of TAO2 as a potential approach for controlling IAV infection. Preliminary data suggest that TAO2
interacts with several splicing factors and other RNA binding proteins. The work outlined in this proposal
seeks a comprehensive understanding of the nuclear activities of TAO2 in human cells and the functional
consequence of these activities for host and viral RNA expression. Specifically, a three-pronged approach
will be taken to: (1) characterize functional TAO2 interaction partners and substrates of phosphorylation
amongst nuclear proteins and determine if IAV infection alters these interactions or if TAO2 interacts
directly with any IAV-encoded proteins; (2) define the role of TAO2 in maintaining the integrity of nuclear
speckles and (3) characterize the global impact of TAO2 on the human splicing machinery and the
consequence of this function for alternative splicing. These goals will be achieved through a combination
of genetic manipulation of cells, biochemistry, mass spectrometry and microscopy. Importantly, the
conclusions obtained from these studies will have implications for general mechanisms of RNA splicing
and nuclear speckle formation and function and will further inform the understanding of host requirements
and vulnerabilities for influenza infection.
Terms: <ATP-protein phosphotransferase><Alternate Splicing><Alternative RNA Splicing><Alternative Splicing><Biochemistry><Biological Chemistry><Cell Body><Cells><Chemicals><Data><Gene Transcription><Genetic Transcription><Goals><Host Factor><Host Factor Protein><Human><Impairment><Influenza Virus><Integration Host Factors><Kinase Family Gene><Kinases><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Messenger RNA><Microscopy><Modern Man><Non-Polyadenylated RNA><Nuclear><Nuclear Proteins><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Pre-mRNA><Protein Kinase><Protein Phosphorylation><Proteins><RNA><RNA Expression><RNA Gene Products><RNA Splicing><RNA, Messenger, Precursors><RNA-Binding Proteins><Regulation><Ribonucleic Acid><Role><Splicing><Transcription><Transphosphorylases><Viral Diseases><Virus Diseases><Virus Replication><Work><flu infection><flu virus infection><gene manipulation><genetic manipulation><genetically manipulate><genetically perturb><glycogen synthase a kinase><hydroxyalkyl protein kinase><infected with flu><infected with flu virus><infected with influenza><infected with influenza virus><influenza infection><influenza virus infection><influenzavirus><mRNA><mRNA Precursor><parent grant><phosphorylase b kinase kinase><social role><trafficking><viral RNA><viral infection><viral multiplication><viral replication><virus RNA><virus infection><virus multiplication><virus-induced disease>