Recent technological advances showed that m6A methylation is abundant in human mRNA and long non- coding RNA and can guide a wide range of cellular processes. The HIV genome comprises numerous putative m6A modification sites and our preliminary experimental data show that m6A modifications are present in the HIV genome. The overall goal of the proposed research is to pinpoint the precise sites of m6Amodifications within the HIV genomic RNA and to determine the functional impact of these modifications on HIV biology.
In Aim 1 we will apply Site-specific Cleavage And Radioactive-labeling followed by Ligation-assisted Extraction and Thin-layer chromatography (SCARLET), to quantify and precisely locate m6A modifications at a single nucleotide resolution.
In Aim 2 we will study the effects of m6A RNA modifications on HIV infectivity and viral production. In addition, we will test the effect of varying m6A in HIV on the immunogenicity of HIV by infecting monocyte derived dendritic cells. The proposed experiments will characterize a novel mode of RNA modification of HIV RNA. The characterization of HIV m6A modifications and its elucidation of its role in the HIV life cycle would not only expand our knowledge of HIV, but also for other viruses that are m6A modified.

Public Health Relevance

We will investigate the effect of N6-methyladenosine (m6A) modifications of the HIV genome on HIV replication and immunogenicity. Our studies of m6A modifications may provide targets for new therapeutic interventions for HIV treatment by either interfering directly with viral replication or by allowing immune recognition.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI125236-01
Application #
9151914
Study Section
Special Emphasis Panel (ZRG1-AARR-P (02)M)
Program Officer
Kuo, Lillian S
Project Start
2016-06-25
Project End
2018-05-31
Budget Start
2016-06-25
Budget End
2017-05-31
Support Year
1
Fiscal Year
2016
Total Cost
$261,882
Indirect Cost
$73,844
Name
Icahn School of Medicine at Mount Sinai
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
Janssens, Sylvie; Schotsaert, Michael; Manganaro, Lara et al. (2018) FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures. Curr Protoc Stem Cell Biol :e65