Site-Specific DNA Cleavage at Orit of the F Plasmid
{'Code': '08070100', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'MCB', 'LongName': 'Div Of Molecular and Cellular Bioscience'}}
{'SignBlockName': 'Philip Harriman', 'PO_EMAI': None, 'PO_PHON': None}
This project will explore the biochemistry of DNA processing at                 the transfer origin (oriT) of the F plasmid from Escherichia coli               K-12.  DNA strand cleavage at oriT is a crucial initial step in                 the conjugal transfer of the F plasmid, which is related to a                   variety of conjugative drug resistance plasmids.  Specific                      cleavage at oriT can occur in the absence of conjugation with the               aid of two to four tra gene products.  The genetic, physical, and               biochemical basis for site-specific strand cleavage at oriT will                be examined.  Oligonucleotide directed mutagenesis will be used                 to identify functional sub-domains of oriT.  Mapping of relevant                tra protein binding sites (e.g., for traI and traY gene products)               will be performed by using extracts and exonuclease III                         protection, and by using purified gene products and footprinting.               DNA/protein complexes ("relaxation complexes") will be sought,                  and their composition and biochemical properties will be                        investigated.  Finally, attempts will be made to reconstitute the               oriT cleavage system in vitro, and to study any higher order                    quaternary complexes by electron microscopy.  These studies may                 help to define in general terms the structural rules for                        DNA/protein interactions and for formation of nucleoprotein                     structures.                                                                                                                                                     The presence of the F factor, a circular DNA plasmid, confers                   upon E. coli the ability to transfer DNA to recipient cells that                lack an F factor.  This process, bacterial conjugation, is                      initiated in the donor by introduction of a break at a specific                 site within one strand of the double-stranded F factor DNA.  One                of the ends created by the break is then transferred into the                   recipient cell.  Dr. Deonier will investigate the biochemical                   mechanism that is responsible for introduction of the break and                 initiation of DNA transfer.  Determination of the exact location                of the break and identifying the plasmid encoded gene products                  that produce it will be major objectives.  This project has the                 potential to provide information on the poorly understood early                 steps in genetic recombination.                                                                                                                                                                                                                                                                                                                                                                                                                                                                           //
{'FirstName': 'Richard', 'LastName': 'Deonier', 'PI_MID_INIT': 'C', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Richard C Deonier', 'EmailAddress': 'deonier@usc.edu', 'NSF_ID': '000362398', 'StartDate': '05/01/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'University of Southern California', 'CityName': 'LOS ANGELES', 'ZipCode': '90033', 'PhoneNumber': '2137407762', 'StreetAddress': '3720 S FLOWER ST FL 3', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'California', 'StateCode': 'CA', 'CONGRESSDISTRICT': '34', 'CONGRESS_DISTRICT_ORG': 'CA34', 'ORG_UEI_NUM': 'G88KLJR3KYT5', 'ORG_LGL_BUS_NAME': 'UNIVERSITY OF SOUTHERN CALIFORNIA', 'ORG_PRNT_UEI_NUM': None}
{'Name': None, 'CityName': None, 'StateCode': None, 'ZipCode': None, 'StreetAddress': None, 'CountryCode': None, 'CountryName': None, 'StateName': None, 'CountryFlag': None, 'CONGRESSDISTRICT': None, 'CONGRESS_DISTRICT_PERF': '""'}
{'Code': '111200', 'Text': 'Genetic Mechanisms'}
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1987&All=true', 'xml': '8717057.xml'}