Understanding The Structural Basis of APOBEC Functions

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DESCRIPTION (provided by applicant): The Structural Basis of APOBEC Functions The APOBEC (Apolioprotein B mRNA-editing Enzyme Catalytic polypeptide) family of cytidine deaminases is a family of enzymes that deaminate cytidine residues of DNA/RNA. APOBEC proteins possess significant cellular functions and anti-viral activity, which partially accounts for the intense attention to this the field of research in recent years. APOBEC proteins are found only in vertebrates and APOBEC3 (Apo3) proteins are found only in primates. By binding and deaminating DNA/RNA, APOBEC enzymes achieve remarkably diverse cellular functions. For example, APOBEC1 (Apo1) edits the mRNA of a protein involved in lipid metabolism; AID plays a key role in somatic hypermutation for antibody maturation; APOBEC2 (Apo2) may play a regulatory role for heart muscle development; and Apo3 proteins, especially Apo3G, can restrict important viral pathogens, including Human Immunodeficiency Virus (HIV) and Hepatitis B Virus (HBV), and retro-element mobility. As a result, a novel approach to HIV therapy focuses on utilizing the potent anti-viral activity of Apo3G and Apo3F. Our long-term goals are to understand the structural/functional relationship for APOBEC cellular function and their anti-viral activity. Our specific aims are to extend our prior success in the structural characterization of APOBEC proteins to the studies of the structural basis of APOBEC's functional mechanisms, including their antiviral activity, with particular focuses on Apo3G and Apo3F. The research will provide valuable information for understanding the molecular details of the APOBEC enzyme family and for the potential drug development to provide therapy for HIV, immune diseases and other diseases related to APOBEC function or malfunction. PUBLIC HEALTH RELEVANCE: Understanding The Structural Basis of APOBEC Functions The Apolioprotein B mRNA-editing enzyme catalytic polypeptide (APOBEC) family of cytidine deaminases are found exclusively in verterbrates. APOBEC nucleic acid deaminases modify genes by deaminating cytosines in mRNA coding sequences and in ssDNA. Their critical biological roles include lipid metabolism, humoral immune response, and potential regulations of developmental process of certain human organs and reproductive system. Additionally, these enzymes can inhibit the replication of retroviruses, such as the human immunodeficiency virus (HIV) and hepatitis B virus (HBV), and retrotransposons. The important beneficial mutational ability of APOBEC proteins can become detrimental to the stability of genome if their activity is not tightly regulated. The understanding of the structural basis of the molecular mechanisms of APOBEC function, which is still poorly understood, bears scientific significance and direct health relevance. We propose to study the structure/function of this important APOBEC deaminase family, with focuses on APOBEC3G and 3F (Apo3G and 3F) proteins and their interactions with cellular and viral ligands, using mainly structural biology, assisted by biophysics, molecular biology, and functional biochemistry.