dna structure modulates the oligomerization properties of the aav initiator protein rep68dna结构调节寡聚化aav发起者蛋白质rep68的属性.pdfVIP

DNA Structure Modulates the Oligomerization Properties of the AAV Initiator Protein Rep68 1 1 2 3 3,4 Jorge Mansilla-Soto , Miran Yoon-Robarts , William J. Rice , Shailee Arya , Carlos R. Escalante *, R. Michael Linden1,5* 1 Department of Gene Cell Medicine, Mount Sinai School of Medicine, New York, New York, United States of America, 2 New York Structural Biology Center, New York, New York, United States of America, 3 Department of Structural and Chemical Biology, Mount Sinai School of Medicine, New York, New York, United States of America, 4 Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond, Virginia, United States of America, 5 Department of Infectious Diseases, King’s College London School of Medicine, Guy’s, King’s College and St. Thomas’ Hospitals, London, United Kingdom Abstract Rep68 is a multifunctional protein of the adeno-associated virus (AAV), a parvovirus that is mostly known for its promise as a gene therapy vector. In addition to its role as initiator in viral DNA replication, Rep68 is essential for site-specific integration of the AAV genome into human chromosome 19. Rep68 is a member of the superfamily 3 (SF3) helicases, along with the well-studied initiator proteins simian virus 40 large T antigen (SV40-LTag) and bovine papillomavirus (BPV) E1. Structurally, SF3 helicases share two domains, a DNA origin interaction domain (OID) and an AAA+ motor domain. The AAA+ motor domain is also a structural feature of cellular initiators and it functions as a platform for initiator oligomerization. Here, we studied Rep68 oligomerization in vitro in the presence of different DNA substrates using a va