Genetic analyses of Schizosaccharomyces pombe dna2(+) reveal that Dna2 plays an essential role in Okazaki fragment metabolism

In this report, we investigated the phenotypes caused by temperature-sensitive (ts) mutant alleles of dna(2+) of Schizosaccharomyces pombe, a homologue of DNA2 of budding yeast, in an attempt to further define its function in vivo with respect to lagging-strand synthesis during the S-phase of the cell cycle. At the restrictive temperature, dna2 (ts) cells arrested at late S-phase but were unaffected in bulk DNA synthesis. Moreover, they exhibited aberrant mitosis when combined xith checkpoint mutations, in keeping with a role for Dna2 in Okazaki fragment maturation. Similarly, spores in which dna2(+) was disrupted duplicated their DNA content during germination and also arrested at late S-phase. Inactivation of dna(2+) led to chromosome fragmentation strikingly similar to that seen when cdc17(+), the DNA ligase I gene, is inactivated The temperature-dependent lethality of dna2 (ts) mutants was suppressed by overexpression of genes encoding subunits of polymerase delta (cdc1(+) and cdc27(+)), DNA ligase I (cdc17(+)), and Fen-l (rad2(+)). Each of these gene products plays a role in the elongation or maturation of Okazaki fragments. Moreover, they all interacted with S. pombe Dna2 in a yeast two-hybrid assay, albeit to different extents. On the basis of these results, ive conclude that dna2(+) plays a direct role in the Okazaki fragment elongation and maturation. We propose that dna2(+) acts as a central protein to form a complex with other proteins required to coordinate the multienzyme process for Okazaki fragment elongation and maturation.
Publisher
GENETICS
Issue Date
2000-07
Language
ENG
Keywords

CELL NUCLEAR ANTIGEN; BASE EXCISION-REPAIR; HIGH-EFFICIENCY TRANSFORMATION; SINGLE-STRANDED-DNA; SACCHAROMYCES-CEREVISIAE; FISSION YEAST; REPLICATION FORK; POLYMERASE-DELTA; LIGASE-I; ENDONUCLEASE ACTIVITY

Citation

GENETICS, v.155, no.3, pp.1055 - 1067

ISSN
0016-6731
URI
http://hdl.handle.net/10203/74838
Appears in Collection
BS-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.
  • Hit : 155
  • Download : 0
  • Cited 0 times in thomson ci
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡClick to seewebofscience_button
⊙ Cited 69 items in WoSClick to see citing articles inrecords_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0