Yeast Mre11 and Rad1 Proteins Define a Ku-Independent Mechanism to Repair Double-Strand Breaks Lacking Overlapping End Sequences

Jia Lin Ma, Eun Mi Kim, James E. Haber, Sang Lee

Research output: Contribution to journalArticle

238 Citations (Scopus)

Abstract

End joining of double-strand breaks (DSBs) requires Ku proteins and frequently involves base pairing between complementary terminal sequences. To define the role of terminal base pairing in end joining, two oppositely oriented HO endonuclease cleavage sites separated by 2.0 kb were integrated into yeast chromosome III, where constitutive expression of HO endonuclease creates two simultaneous DSBs with no complementary end sequence. Lack of complementary sequence in their 3′ single-strand overhangs facilitates efficient repair events distinctly different from when the 3′ ends have a 4-bp sequence base paired in various ways to create 2- to 3-bp insertions. Repair of noncomplementary ends results in a set of nonrandom deletions of up to 302 bp, annealed by imperfect microhomology of about 8 to 10 bp at the junctions. This microhomology-mediated end joining (MMEJ) is Ku independent, but strongly dependent on Mre11, Rad50, and Rad1 proteins and partially dependent on Dnl4 protein. The MMEJ also occurs when Rad52 is absent, but the extent of deletions becomes more limited. The increased gamma ray sensitivity of rad1Δ rad52Δ yku70Δ strains compared to rad52Δ yku70Δ strains suggests that MMEJ also contributes to the repair of DSBs induced by ionizing radiation.

Original languageEnglish (US)
Pages (from-to)8820-8828
Number of pages9
JournalMolecular and Cellular Biology
Volume23
Issue number23
DOIs
StatePublished - Dec 2003

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Endonucleases
Base Pairing
Yeasts
Gamma Rays
Ionizing Radiation
Proteins
Chromosomes
Ku Autoantigen

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Yeast Mre11 and Rad1 Proteins Define a Ku-Independent Mechanism to Repair Double-Strand Breaks Lacking Overlapping End Sequences. / Ma, Jia Lin; Kim, Eun Mi; Haber, James E.; Lee, Sang.

In: Molecular and Cellular Biology, Vol. 23, No. 23, 12.2003, p. 8820-8828.

Research output: Contribution to journalArticle

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