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Gene Review

RFA2  -  Rfa2p

Saccharomyces cerevisiae S288c

Synonyms: BUF1, DNA-binding protein BUF1, N0368, RF-A protein 2, RPA2, ...
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High impact information on RFA2

  • A truncation of the N-terminal region of Rfa2p (associated with the rfa2Delta40 mutated allele) results in severe telomere shortening caused by a defect in the in vivo regulation of telomerase activity [1].
  • Our results correlate the set1Delta-dependent phosphorylation of Rfa2p with the transcriptional induction of repair genes [2].
  • This work provides an additional link for the role of Rfa2p in the regulation of the repair capacity of the cell and reveals a role for the phosphorylation of Rfa2p and unveils unsuspected connections between chromatin, signaling pathways, telomeres, and DNA repair [2].
  • Replication factor-A (RF-A) is a three-subunit protein complex originally purified from human cells as an essential component for SV40 DNA replication in vitro [3].
  • RF-A from both human and yeast cells is phosphorylated in a cell-cycle-dependent manner; the protein is phosphorylated at the G1- to S-phase transition, and dephosphorylation occurs at mitosis, thereby resetting this cycle [4].

Biological context of RFA2

  • We demonstrated that null alleles of DUN1 and certain mutant alleles of RFA2 result in short telomeres [5].
  • As observed with Xrs2p, however, strains with mutations of DUN1 or RFA2 that eliminate SQ motifs have no effect on telomere length or DNA damage sensitivity [5].
  • Amino acid changes in Xrs2p, Dun1p, and Rfa2p that remove the preferred targets of the ATM family of protein kinases do not affect DNA repair or telomere length in Saccharomyces cerevisiae [5].
  • Replication protein A (RPA), the heterotrimeric single-stranded-DNA (ssDNA) binding protein (SSB) of eukaryotes, contains two homologous ssDNA binding domains (A and B) in its largest subunit, RPA1, and a third domain in its second-largest subunit, RPA2 [6].
  • These results suggest an essential role of RPA2 in the maintenance of transcriptional gene silencing at specific loci in a DNA-methylation-independent manner [7].

Associations of RFA2 with chemical compounds

  • Finally, rfa2 mutant cells have a mutator and hyper-recombination phenotype and are more sensitive to hydroxyurea and methyl-methane-sulfonate than wild-type cells [8].

Regulatory relationships of RFA2


Other interactions of RFA2

  • To identify proteins interacting with the Mec1-Ddc2 complex, we performed a modified two-hybrid screen and isolated RFA1 and RFA2, genes that encode subunits of replication protein A (RPA) [10].
  • The nuclear import of the RPA complex subunit Rfa2 is impaired in nup82-3 and in mutants of the karyopherin KAP95, but is not affected by the loss of MSN5 [11].
  • Moreover, when shifted to the restrictive temperature, about 50% of the rfa2 mutant cells rapidly die while traversing the S phase and the surviving cells arrest in late S/G2 at the RAD9 checkpoint [8].
  • The in vivo function of the 34 kDa subunit of yeast replication protein A (RPA), encoded by the RFA2 gene, has been studied by analyzing the effect of Rpa34 depletion and by producing and characterizing rfa2 temperature-sensitive mutants [8].
  • Primary Rfa2 phosphorylation occurs early in meiotic progression and is independent of DNA replication, recombination and Mec1 [9].


  1. RPA regulates telomerase action by providing Est1p access to chromosome ends. Schramke, V., Luciano, P., Brevet, V., Guillot, S., Corda, Y., Longhese, M.P., Gilson, E., Géli, V. Nat. Genet. (2004) [Pubmed]
  2. The set1Delta mutation unveils a novel signaling pathway relayed by the Rad53-dependent hyperphosphorylation of replication protein A that leads to transcriptional activation of repair genes. Schramke, V., Neecke, H., Brevet, V., Corda, Y., Lucchini, G., Longhese, M.P., Gilson, E., Géli, V. Genes Dev. (2001) [Pubmed]
  3. Replication factor-A from Saccharomyces cerevisiae is encoded by three essential genes coordinately expressed at S phase. Brill, S.J., Stillman, B. Genes Dev. (1991) [Pubmed]
  4. Cell-cycle-regulated phosphorylation of DNA replication factor A from human and yeast cells. Din, S., Brill, S.J., Fairman, M.P., Stillman, B. Genes Dev. (1990) [Pubmed]
  5. Amino acid changes in Xrs2p, Dun1p, and Rfa2p that remove the preferred targets of the ATM family of protein kinases do not affect DNA repair or telomere length in Saccharomyces cerevisiae. Mallory, J.C., Bashkirov, V.I., Trujillo, K.M., Solinger, J.A., Dominska, M., Sung, P., Heyer, W.D., Petes, T.D. DNA Repair (Amst.) (2003) [Pubmed]
  6. Identification and characterization of the fourth single-stranded-DNA binding domain of replication protein A. Brill, S.J., Bastin-Shanower, S. Mol. Cell. Biol. (1998) [Pubmed]
  7. Mutations in a conserved replication protein suppress transcriptional gene silencing in a DNA-methylation-independent manner in Arabidopsis. Kapoor, A., Agarwal, M., Andreucci, A., Zheng, X., Gong, Z., Hasegawa, P.M., Bressan, R.A., Zhu, J.K. Curr. Biol. (2005) [Pubmed]
  8. Mutations in the gene encoding the 34 kDa subunit of yeast replication protein A cause defective S phase progression. Santocanale, C., Neecke, H., Longhese, M.P., Lucchini, G., Plevani, P. J. Mol. Biol. (1995) [Pubmed]
  9. Replication protein A is sequentially phosphorylated during meiosis. Brush, G.S., Clifford, D.M., Marinco, S.M., Bartrand, A.J. Nucleic Acids Res. (2001) [Pubmed]
  10. Role of the C terminus of Mec1 checkpoint kinase in its localization to sites of DNA damage. Nakada, D., Hirano, Y., Tanaka, Y., Sugimoto, K. Mol. Biol. Cell (2005) [Pubmed]
  11. The karyopherin Msn5/Kap142 requires Nup82 for nuclear export and performs a function distinct from translocation in RPA protein import. Belanger, K.D., Simmons, L.A., Roth, J.K., VanderPloeg, K.A., Lichten, L.B., Fahrenkrog, B. J. Biol. Chem. (2004) [Pubmed]
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