P38859 (DNA2_YEAST) Reviewed, UniProtKB/Swiss-Prot
Last modified February 19, 2014. Version 120. History...
Names and origin
|Protein names||Recommended name:|
DNA replication ATP-dependent helicase/nuclease DNA2
|Organism||Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast) [Reference proteome]|
|Taxonomic identifier||559292 [NCBI]|
|Taxonomic lineage||Eukaryota › Fungi › Dikarya › Ascomycota › Saccharomycotina › Saccharomycetes › Saccharomycetales › Saccharomycetaceae › Saccharomyces ›|
|Sequence length||1522 AA.|
|Protein existence||Evidence at protein level|
General annotation (Comments)
Key enzyme involved in DNA replication and DNA repair. Involved in Okazaki fragments processing by cleaving long flaps that escape FEN1: flaps that are longer than 27 nucleotides are coated by replication protein A complex (RPA), leading to recruit DNA2 which cleaves the flap until it is too short to bind RPA and becomes a substrate for FEN1. Also involved in 5'-end resection of DNA during double-strand break (DSB) repair by mediating the cleavage of 5'-ssDNA. Possesses different enzymatic activities, such as single-stranded DNA (ssDNA)-dependent ATPase, 5'-3' helicase and endonuclease activities. While the ATPase and endonuclease activities are well-defined and play a key role in Okazaki fragments processing and DSB repair, the 5'-3' DNA helicase activity is atypical: it cannot load onto its tracking strand internally and has an absolute free 5'-end requirement. Helicase activity may promote the motion of DNA2 on the flap, helping the nuclease function. Ref.4 Ref.5 Ref.6 Ref.8 Ref.9 Ref.10
ATP + H2O = ADP + phosphate.
Binds 1 4Fe-4S cluster. Ref.11
Phosphorylated at Ser-17 and Ser-237 by CDK1 in response to DNA damage, leading to promote recruitment to double-strand break (DSB) sites and DNA resection. Ref.10
Belongs to the DNA2/NAM7 helicase family.
Sequence annotation (Features)
|Feature key||Position(s)||Length||Description||Graphical view||Feature identifier|
|Chain||1 – 1522||1522||DNA replication ATP-dependent helicase/nuclease DNA2||PRO_0000080711|
|Nucleotide binding||1074 – 1081||8||ATP By similarity|
|Region||450 – 900||451||Nuclease activity|
|Region||901 – 1522||622||Helicase activity|
|Metal binding||519||1||Iron-sulfur (4Fe-4S)|
|Metal binding||768||1||Iron-sulfur (4Fe-4S)|
|Metal binding||771||1||Iron-sulfur (4Fe-4S)|
|Metal binding||777||1||Iron-sulfur (4Fe-4S)|
Amino acid modifications
|Modified residue||4||1||Phosphothreonine Ref.10|
|Modified residue||17||1||Phosphoserine; by CDK1 Ref.10|
|Modified residue||237||1||Phosphoserine; by CDK1 Ref.10|
|Modified residue||962||1||Phosphothreonine Ref.7|
|Mutagenesis||4||1||T → A: Abolishes phosphorylation by CDK1, leading to a poor recruitment at double-strand. break (DSB) sites following DNA damage; when associated with A-17 and A-237. Ref.10|
|Mutagenesis||17||1||S → A: Abolishes phosphorylation by CDK1, leading to a poor recruitment at double-strand. break (DSB) sites following DNA damage; when associated with A-4 and A-237. Ref.10|
|Mutagenesis||17||1||S → D: Mimics phosphorylation; restores nuclear localization and recruitment at double-strand. break (DSB) sites following DNA damage; when associated with D-237. Ref.10|
|Mutagenesis||237||1||S → A: Abolishes phosphorylation by CDK1, leading to a poor recruitment at double-strand. break (DSB) sites following DNA damage; when associated with A-4 and A-17. Ref.10|
|Mutagenesis||237||1||S → D: Mimics phosphorylation; restores nuclear localization and recruitment at double-strand. break (DSB) sites following DNA damage; when associated with D-17. Ref.10|
|Mutagenesis||504||1||P → S in dna2-1; temperature-sensitive mutant unable to grow at 37 degrees Celsius, probably due to abolition of iron-sulfur-binding. Ref.11|
|Mutagenesis||519||1||C → A: Abolishes iron-sulfur-binding; when associated with A-768; A-771 and A-777. Impaired nuclease and ATPase activities; when associated with A-768. Ref.11|
|Mutagenesis||675||1||E → A: Nuclease dead mutant. No helicase activity when the 5'-end of the substrate is blocked. Ref.8|
|Mutagenesis||677||1||K → R: Nuclease dead mutant. No helicase activity when the 5'-end of the substrate is blocked. Ref.8|
|Mutagenesis||768||1||C → A: Abolishes iron-sulfur-binding; when associated with A-519; A-771 and A-777. Impaired nuclease and ATPase activities; when associated with A-519. Ref.11|
|Mutagenesis||771||1||C → A: Abolishes iron-sulfur-binding; when associated with A-519; A-768 and A-777. Impaired nuclease and ATPase activities; when associated with A-777. Ref.11|
|Mutagenesis||777||1||C → A: Abolishes iron-sulfur-binding; when associated with A-519; A-768 and A-771. Impaired nuclease and ATPase activities; when associated with A-771. Ref.11|
|||"Complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII."|
Johnston M., Andrews S., Brinkman R., Cooper J., Ding H., Dover J., Du Z., Favello A., Fulton L., Gattung S., Geisel C., Kirsten J., Kucaba T., Hillier L.W., Jier M., Johnston L., Langston Y., Latreille P. Vaudin M.
Science 265:2077-2082(1994) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: ATCC 204508 / S288c.
|||Saccharomyces Genome Database|
Submitted (DEC-2009) to the EMBL/GenBank/DDBJ databases
Cited for: GENOME REANNOTATION.
Strain: ATCC 204508 / S288c.
|||"A yeast gene required for DNA replication encodes a protein with homology to DNA helicases."|
Budd M.E., Campbell J.L.
Proc. Natl. Acad. Sci. U.S.A. 92:7642-7646(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: CHARACTERIZATION.
|||"Dna2 of Saccharomyces cerevisiae possesses a single-stranded DNA-specific endonuclease activity that is able to act on double-stranded DNA in the presence of ATP."|
Bae S.H., Choi E., Lee K.H., Park J.S., Lee S.H., Seo Y.S.
J. Biol. Chem. 273:26880-26890(1998) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
|||"Dna2p helicase/nuclease is a tracking protein, like FEN1, for flap cleavage during Okazaki fragment maturation."|
Kao H.I., Campbell J.L., Bambara R.A.
J. Biol. Chem. 279:50840-50849(2004) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
|||"Sgs1 helicase and two nucleases Dna2 and Exo1 resect DNA double-strand break ends."|
Zhu Z., Chung W.H., Shim E.Y., Lee S.E., Ira G.
Cell 134:981-994(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
|||"A multidimensional chromatography technology for in-depth phosphoproteome analysis."|
Albuquerque C.P., Smolka M.B., Payne S.H., Bafna V., Eng J., Zhou H.
Mol. Cell. Proteomics 7:1389-1396(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT THR-962, MASS SPECTROMETRY.
|||"Dna2 exhibits a unique strand end-dependent helicase function."|
Balakrishnan L., Polaczek P., Pokharel S., Campbell J.L., Bambara R.A.
J. Biol. Chem. 285:38861-38868(2010) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF GLU-675 AND LYS-677.
|||"DNA end resection by Dna2-Sgs1-RPA and its stimulation by Top3-Rmi1 and Mre11-Rad50-Xrs2."|
Cejka P., Cannavo E., Polaczek P., Masuda-Sasa T., Pokharel S., Campbell J.L., Kowalczykowski S.C.
Nature 467:112-116(2010) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
|||"Cell cycle regulation of DNA double-strand break end resection by Cdk1-dependent Dna2 phosphorylation."|
Chen X., Niu H., Chung W.H., Zhu Z., Papusha A., Shim E.Y., Lee S.E., Sung P., Ira G.
Nat. Struct. Mol. Biol. 18:1015-1019(2011) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, PHOSPHORYLATION AT THR-4; SER-17 AND SER-237, MUTAGENESIS OF THR-4; SER-17 AND SER-237.
|||"Cross talk between the nuclease and helicase activities of Dna2: role of an essential iron-sulfur cluster domain."|
Pokharel S., Campbell J.L.
Nucleic Acids Res. 40:7821-7830(2012) [PubMed] [Europe PMC] [Abstract]
Cited for: COFACTOR, IRON-SULFUR-BINDING, MUTAGENESIS OF PRO-504; CYS-519; CYS-768; CYS-771 AND CYS-777.
|+||Additional computationally mapped references.|
|U00027 Genomic DNA. Translation: AAB68010.1.|
BK006934 Genomic DNA. Translation: DAA06857.1.
|RefSeq||NP_012034.1. NM_001179295.1. |
3D structure databases
|SMR||P38859. Positions 1049-1466. |
Protein-protein interaction databases
|BioGrid||36598. 76 interactions.|
|IntAct||P38859. 8 interactions.|
Protocols and materials databases
Genome annotation databases
|EnsemblFungi||YHR164C; YHR164C; YHR164C. |
|SGD||S000001207. DNA2. |
Enzyme and pathway databases
|Reactome||REACT_160098. Cytosolic Iron-sulfur Cluster Assembly (yeast). |
Gene expression databases
Family and domain databases
|InterPro||IPR026851. Dna2. |
|PANTHER||PTHR10887:SF14. PTHR10887:SF14. 1 hit. |
|Pfam||PF08696. Dna2. 1 hit. |
|SUPFAM||SSF52540. SSF52540. 2 hits. |
|Accession||Primary (citable) accession number: P38859|
Secondary accession number(s): D3DLB3
|Entry status||Reviewed (UniProtKB/Swiss-Prot)|
|Annotation program||Fungal Protein Annotation Program|