Abstract

A 120 nucleotide bacteriophage PHI29 RNA is structural and catalytic constituent of the viral precursor shell (prohead) into which the 18 kilobase pair double-stranded viral DNA is packaged in an efficient, ATP-dependent, in vitro reaction. The RNA is a part of the portal vertex of the prohead (connector), which together with the PHI29 DNA packaging protein gp16 may function as an ATPase needed for translocation of DNA-gp3 (PHI29 DNA has a covalently bound protein gp3 at each 5' terminus). Thus the PHI29 DNA """"""""packaging machine"""""""" includes the connector protein (gp10), RNA, gp16, and perhaps the gp3 of DNA-gp3. The role of the RNA in the sequential initiation and translocation events of in vitro DNA-gp3 packaging and its possible function as part of the ATPase will be investigated. Localization of the RNA in the connector will be attempted by processing of electron micrographs and three-dimensional reconstructions. Prohead RNAs of PHI29 relatives and other phages will be isolated and sequenced. The higher-order structure of PHI29 prohead RNA and counterpart RNAs will be studied by the use of comparative sequence analysis, footprinting, and chemical and enzymatic probes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM039931-03
Application #
3297227
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1988-04-01
Project End
1993-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
University of Minnesota Twin Cities
Department
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455

  Publications

Zhang, F; Lemieux, S; Wu, X et al. (1998) Function of hexameric RNA in packaging of bacteriophage phi 29 DNA in vitro. Mol Cell 2:141-7
Zhang, F; Anderson, D (1998) In vitro selection of bacteriophage phi29 prohead RNA aptamers for prohead binding. J Biol Chem 273:2947-53
Tsuprun, V; Anderson, D; Egelman, E H (1994) The bacteriophage phi 29 head-tail connector shows 13-fold symmetry in both hexagonally packed arrays and as single particles. Biophys J 66:2139-50
Wichitwechkarn, J; Johnson, D; Anderson, D (1992) Mutant prohead RNAs in the in vitro packaging of bacteriophage phi 29 DNA-gp3. J Mol Biol 223:991-8
Grimes, S; Anderson, D (1990) RNA dependence of the bacteriophage phi 29 DNA packaging ATPase. J Mol Biol 215:559-66
Anderson, D; Bodley, J W (1990) Role of RNA in bacteriophage phi 29 DNA packaging. J Struct Biol 104:70-4
Grimes, S; Anderson, D (1989) In vitro packaging of bacteriophage phi 29 DNA restriction fragments and the role of the terminal protein gp3. J Mol Biol 209:91-100
Wichitwechkarn, J; Bailey, S; Bodley, J W et al. (1989) Prohead RNA of bacteriophage phi 29: size, stoichiometry and biological activity. Nucleic Acids Res 17:3459-68
Sen, P; Pazour, G J; Anderson, D et al. (1989) Cooperative binding of Agrobacterium tumefaciens VirE2 protein to single-stranded DNA. J Bacteriol 171:2573-80