Project Grant F31AI202950

THE HUMAN INTEGRATOR COMPLEX IS REQUIRED FOR HERPES SIMPLEX VIRUS TYPE 1 INFECTION - ABSTRACT HERPES SIMPLEX VIRUS TYPE 1 (HSV-1) IS A WIDESPREAD, DOUBLE-STRANDED DNA ALPHA HERPESVIRUS BEST KNOWN FOR ITS CAPACITY TO ESTABLISH LIFELONG LATENCY WITHIN NEURONS. ALTHOUGH MANY INFECTED INDIVIDUALS EXPERIENCE NO SYMPTOMS OR ONLY RECURRENT ORAL LESIONS, HSV-1 REMAINS THE PRIMARY CAUSE OF FATAL VIRAL ENCEPHALITIS AND IS A MAJOR CONTRIBUTOR TO INFECTIOUS BLINDNESS. DESPITE ITS SUBSTANTIAL GLOBAL BURDEN, NO LICENSED VACCINE AGAINST HSV- 1 EXISTS AND ANTIVIRAL AGENTS SUCH AS ACYCLOVIR PROVIDE ONLY LIMITED BENEFIT UNLESS ADMINISTERED EARLY IN THE LYTIC PHASE. HSV-1 DEPENDS ENTIRELY ON HOST RNA POLYMERASE II (RNAPII) TO DRIVE VIRAL GENE EXPRESSION, YET THE MOLECULAR MECHANISMS THAT COORDINATE INTERACTIONS BETWEEN VIRAL FACTORS AND HOST TRANSCRIPTIONAL REGULATORS REMAIN POORLY DEFINED. THIS PROPOSAL SEEKS TO CLOSE THIS CRITICAL GAP BY EXAMINING HOW THE HOST RNAPII REGULATORY APPARATUS IS MODULATED DURING HSV-1 INFECTION. THE INTEGRATOR COMPLEX (INT) IS A MULTI-SUBUNIT, MODULAR ASSEMBLY THAT ENGAGES PAUSED RNAPII AND FACILITATES PROMOTER-PROXIMAL TRANSCRIPTION TERMINATION. IN DOING SO, INT SUPPRESSES EXPRESSION OF PROTEIN-CODING GENES AND GOVERNS 3-END PROCESSING OF SHORT NONCODING TRANSCRIPTS, INCLUDING ENHANCER RNAS (ERNAS). WORK FROM THE WAGNER LABORATORY AND OTHERS HAS DEMONSTRATED THAT INT CARRIES OUT RNAPII TERMINATION VIA TWO SEPARABLE ENZYMATIC FUNCTIONS: AN RNA ENDONUCLEASE ACTIVITY RESIDING IN INTEGRATOR SUBUNIT 11 (INTS11) AND A PHOSPHATASE ACTIVITY MEDIATED BY PROTEIN PHOSPHATASE 2A (PP2A), WHICH DIRECTLY ASSOCIATES WITH INTS8. ALTHOUGH INT IS ESTABLISHED AS A CENTRAL REGULATOR OF RNAPII DYNAMICS, ITS CONTRIBUTION TO HSV-1 INFECTION REMAINS LARGELY UNEXPLORED. TO PROBE THIS QUESTION, WE GENERATED AN AUXIN-INDUCIBLE DEGRON (AID) SYSTEM THAT ENABLES RAPID (