RFA2/YNL312W Summary Help

Standard Name RFA2 1
Systematic Name YNL312W
Alias BUF1 , RPA2 , RPA32 2
Feature Type ORF, Verified
Description Subunit of heterotrimeric Replication Protein A (RPA); RPA is a highly conserved single-stranded DNA binding protein involved in DNA replication, repair, and recombination; RPA protects against inappropriate telomere recombination, and upon telomere uncapping, prevents cell proliferation by a checkpoint-independent pathway; in concert with Sgs1p-Top2p-Rmi1p, stimulates DNA catenation/decatenation activity of Top3p; protein abundance increases in response to DNA replication s (1, 3, 4, 5, 6, 7 and see Summary Paragraph)
Name Description Replication Factor A 1
Chromosomal Location
ChrXIV:48287 to 49216 | ORF Map | GBrowse
Gene Ontology Annotations All RFA2 GO evidence and references
  View Computational GO annotations for RFA2
Molecular Function
Manually curated
Biological Process
Manually curated
Cellular Component
Manually curated
Regulators 5 genes
Classical genetics
reduction of function
Large-scale survey
reduction of function
255 total interaction(s) for 176 unique genes/features.
Physical Interactions
  • Affinity Capture-MS: 44
  • Affinity Capture-RNA: 1
  • Affinity Capture-Western: 13
  • Biochemical Activity: 8
  • Co-localization: 1
  • Co-purification: 1
  • Reconstituted Complex: 7
  • Two-hybrid: 11

Genetic Interactions
  • Dosage Lethality: 1
  • Dosage Rescue: 1
  • Negative Genetic: 109
  • Phenotypic Enhancement: 1
  • Positive Genetic: 49
  • Synthetic Growth Defect: 3
  • Synthetic Lethality: 5

Expression Summary
Length (a.a.) 273
Molecular Weight (Da) 29,936
Isoelectric Point (pI) 4.7
Phosphorylation PhosphoGRID | PhosphoPep Database
sequence information
ChrXIV:48287 to 49216 | ORF Map | GBrowse
Last Update Coordinates: 2011-02-03 | Sequence: 1996-07-31
Subfeature details
Most Recent Updates
Coordinates Sequence
CDS 1..7 48287..48293 2011-02-03 1996-07-31
Intron 8..115 48294..48401 2011-02-03 1996-07-31
CDS 116..930 48402..49216 2011-02-03 1996-07-31
Retrieve sequences
Analyze Sequence
S288C only
S288C vs. other species
S288C vs. other strains
External Links All Associated Seq | Entrez Gene | Entrez RefSeq Protein | MIPS | Search all NCBI (Entrez) | UniProtKB
Primary SGDIDS000005256

RFA1, RFA2, and RFA3 encode the three subunits of the heterotrimeric complex Replication protein A (RPA). RPA is a highly conserved single-stranded DNA (ssDNA) binding protein that removes secondary structure from ssDNA. RPA plays a key role in DNA metabolic pathways that have a ssDNA intermediate such as replication, recombination, transcription, checkpoints, telomere maintenance, and DNA repair (reviewed in 8 and 9). Some specific instances of RPA function in yeast include loading the telomerase component Est1p onto telomeres (10), enabling the efficient assembly of the Rad51p presynaptic filament (11, 12), and providing specificity of Rad17p/Mec3p/Ddc1p loading at 5'-junctions (13).

RPA binds ssDNA with high affinity and has no sequence specificity but does preferentially bind to pyrimidines over purines (4, 14). RPA interaction with DNA is mediated by binding domains in Rfa1p and Rfa2p (15). In response to DNA damage, Rfa1p and Rfa2p are phosphorylated by overlapping mechanisms that involve the kinases Mec1p and Tel1p (16, 17, 18). These same kinases also mediate Rfa2p phosphorylation during the normal cell cycle and during meiosis (16, 19).

All three RFA subunits are essential for viability in yeast, but reduction of function mutations in RPA result in various defects such as sensitivity to DNA damaging agents, altered mitotic and meiotic recombination, failure in mating type switching, severe telomere shortening, and chromosomal instability (10, 12 and reviewed in 9, 20). Gross chromosomal rearrangements found in rfa1 mutants are similar to those seen in some human cancers, and corresponding mutations in mice lead to tumor development (21 and references therein).

Last updated: 2007-08-09 Contact SGD

References cited on this page View Complete Literature Guide for RFA2
1) Brill SJ and Stillman B  (1991) Replication factor-A from Saccharomyces cerevisiae is encoded by three essential genes coordinately expressed at S phase. Genes Dev 5(9):1589-600
2) Hegnauer AM, et al.  (2012) An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. EMBO J 31(18):3768-83
3) Longhese MP, et al.  (1994) Replication factor A is required in vivo for DNA replication, repair, and recombination. Mol Cell Biol 14(12):7884-90
4) Alani E, et al.  (1992) Characterization of DNA-binding and strand-exchange stimulation properties of y-RPA, a yeast single-strand-DNA-binding protein. J Mol Biol 227(1):54-71
5) Tkach JM, et al.  (2012) Dissecting DNA damage response pathways by analysing protein localization and abundance changes during DNA replication stress. Nat Cell Biol 14(9):966-76
6) Cejka P, et al.  (2012) Decatenation of DNA by the S. cerevisiae Sgs1-Top3-Rmi1 and RPA complex: a mechanism for disentangling chromosomes. Mol Cell 47(6):886-96
7) Grandin N and Charbonneau M  (2013) RPA provides checkpoint-independent cell cycle arrest and prevents recombination at uncapped telomeres of Saccharomyces cerevisiae. DNA Repair (Amst) 12(3):212-26
8) Sung P, et al.  (2003) Rad51 recombinase and recombination mediators. J Biol Chem 278(44):42729-32
9) Symington LS  (2002) Role of RAD52 epistasis group genes in homologous recombination and double-strand break repair. Microbiol Mol Biol Rev 66(4):630-70, table of contents
10) Schramke V, et al.  (2004) RPA regulates telomerase action by providing Est1p access to chromosome ends. Nat Genet 36(1):46-54
11) Hays SL, et al.  (1998) Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA. Mol Cell Biol 18(7):4400-6
12) Kantake N, et al.  (2003) The recombination-deficient mutant RPA (rfa1-t11) is displaced slowly from single-stranded DNA by Rad51 protein. J Biol Chem 278(26):23410-7
13) Majka J, et al.  (2006) Replication protein A directs loading of the DNA damage checkpoint clamp to 5'-DNA junctions. J Biol Chem 281(38):27855-61
14) Kim C, et al.  (1992) Binding properties of replication protein A from human and yeast cells. Mol Cell Biol 12(7):3050-9
15) Brill SJ and Bastin-Shanower S  (1998) Identification and characterization of the fourth single-stranded-DNA binding domain of replication protein A. Mol Cell Biol 18(12):7225-34
16) Brush GS, et al.  (1996) The ATM homologue MEC1 is required for phosphorylation of replication protein A in yeast. Proc Natl Acad Sci U S A 93(26):15075-80
17) Brush GS and Kelly TJ  (2000) Phosphorylation of the replication protein A large subunit in the Saccharomyces cerevisiae checkpoint response. Nucleic Acids Res 28(19):3725-32
18) Kim HS and Brill SJ  (2003) MEC1-dependent phosphorylation of yeast RPA1 in vitro. DNA Repair (Amst) 2(12):1321-35
19) Brush GS, et al.  (2001) Replication protein A is sequentially phosphorylated during meiosis. Nucleic Acids Res 29(23):4808-17
20) Sung P, et al.  (2000) Recombination factors of Saccharomyces cerevisiae. Mutat Res 451(1-2):257-75
21) Wang Y, et al.  (2005) Mutation in Rpa1 results in defective DNA double-strand break repair, chromosomal instability and cancer in mice. Nat Genet 37(7):750-5