title: A single zinc finger optimizes the DNA interactions of the nucleocapsid protein of the yeast retrotransposon Ty3.
abstract: Reverse transcription in retroviruses and retrotransposons requires nucleic acid chaperones, which drive the rearrangement of nucleic acid conformation. The nucleic acid chaperone properties of the human immunodeficiency virus type-1 (HIV-1) nucleocapsid (NC) protein have been extensively studied, and nucleic acid aggregation, duplex destabilization and rapid binding kinetics have been identified as major components of its activity. However, the properties of other nucleic acid chaperone proteins, such as retrotransposon Ty3 NC, a likely ancestor of HIV-1 NC, are not well understood. In addition, it is unclear whether a single zinc finger is sufficient to optimize the properties characteristic of HIV-1 NC. We used single-molecule DNA stretching as a method for detailed characterization of Ty3 NC chaperone activity. We found that wild type Ty3 NC aggregates single- and double-stranded DNA, weakly stabilizes dsDNA, and exhibits rapid binding kinetics. Single-molecule studies in the presence of Ty3 NC mutants show that the N-terminal basic residues and the unique zinc finger at the C-terminus are required for optimum chaperone activity in this system. While the single zinc finger is capable of optimizing Ty3 NC's DNA interaction kinetics, two zinc fingers may be necessary in order to facilitate the DNA destabilization exhibited by HIV-1 NC.
authors.affiliation: Department of Physics, Northeastern University, Boston, MA, USA.
authors.forename: Kathy R
authors.initials: KR
authors.lastname: Chaurasiya
authors.forename: Hylkje
authors.initials: H
authors.lastname: Geertsema
authors.forename: Gaël
authors.initials: G
authors.lastname: Cristofari
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Darlix
authors.forename: Mark C
authors.initials: MC
authors.lastname: Williams
mesh_terms: D000595:Amino Acid Sequence; D004247:DNA / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018626:Retroelements; D017384:Sequence Deletion; D015003:Yeasts / Q000235:genetics; D016335:Zinc Fingers
chemical_list: D019590:Nucleocapsid Proteins; D018626:Retroelements; D004247:DNA