Table 1. Summary of the most damaging effects of Rad50 mutations obtained from previous in vitro and in vivo experiments.
| Motif/domain | Mutations | Organism | Effects | References |
|---|---|---|---|---|
| Walker A | K40A/R/E | S. cerevisiae | • HR and NHEJ defects and lower ATPase activity | Chen et al. (2005) |
| Walker A D-loop | N38A, D512N/A | T4 bacteriophage | • Naturally occurring mutation of CFTR protein | De La Rosa & Nelson (2011) |
| • Reduce in ATP activity | ||||
| ATP binding domain and Walker A | G39D, K40E, K81I, R20M | S. cerevisiae | • Total defect in formation of viable spore | Alani, Padmore & Kleckner (1990) |
| ATP binding domain | K6E, K22M, R83I | M. musculus | • Embryonic lethality, growth defect, cancer predisposition, hematopoietic and spermatogenic depletion | Bender et al. (2002) |
| Walker A | K39R, K42M | D. radiodurans | • Prevented ATP binding and hydrolysis | Koroleva et al. (2007) |
| ATPase binding domain, Walker B and Signature motif | K115E, K175E, K182E, R94E, K95E, R765E | T. maritima | In vitro: Thermotoga maritima | Rojowska et al. (2014) |
| • K175E, K182E, K115E Reduced DNA binding | ||||
| • R94E and K95E: Important for DNA binding | ||||
| • R765E: Diminished DNA binding | ||||
| • E798Q: Low affinity to DNA | ||||
| • S768R: Reduced DNA binding | ||||
| E798Q, S768R, K103E, K104E, R131E, R1202E, S1205R, E1235Q | S. cerevisiae | In vivo: Saccharomyces cerevisiae | ||
| • S1205R and E1235Q double mutation: Unable to rescue the impaired DNA damage response | ||||
| • K103E, K104E and R131E: Strongly affected DNA binding and moderate reduction in telomere length | ||||
| • K103E and R131R (double mutation) and R1201E: Significantly reduced telomere length | ||||
| • S1205R: Significantly reduced telomere length | ||||
| Zinc hook | S679R, P682E, V683R | M. musculus | • Lethality in mice. Hydrocephalus, defects in primitive hematopoietic and gametogenic cells | Roset et al. (2014) |
| C684N, C685A, P686A, V6871, C688R, Q689S | S. cerevisiae | • Defective to be recruited to chromosomal double strand break | He et al. (2012) | |
| • Phenotype as severe as Rad50 null mutant | ||||
| • Defective in ATM activation, HR, sensitive to irradiation and ATR activation | ||||
| C288S, C291S | T4 bacteriophage | • Double mutation is lethal | Barfoot et al. (2015) | |
| S635G | H. sapiens | • Chromosomal instability | Gatei et al. (2011) | |
| • Defective ATM-dependent signaling | ||||
| S685R, Y688E, L689R | S. cerevisiae | • S685R and Y688E double mutation: Sporulation efficiency and viability were severely impaired followed by L689R | Hohl et al. (2015) | |
| • Rad50-Mre11 interaction was strongly impaired, partial suppression of telomere and meiotic defects | ||||
| Rad50 Signature motif | R805E, L802W | P. furiosus | • L802W: Decrease dimerization in ATP, hydrolysis and cleavage site | Deshpande et al. (2014) |
| • R805E: Poorly grown in camptothecin; inability to repair endogenous DNA damage by HR and showed defect in resection in HO endonuclease induced | ||||
| K1187A, K1187E, R1195A, R1195E | S. pombe | • K1187A: Sensitive in higher dose of clastogens | Williams et al. (2011) | |
| • K1187E, R1195A and R1195E: Significantly sensitive to clastogen agents and were deleterious as Rad50 null mutation | ||||
| S471A/R/M, E474Q, K475M | T4 bacteriophage | • S471A/R.M, E474Q and K475M: Residues involved in the allosteric transmission between DNA and ATP binding sites | Herdendorf & Nelson (2011) | |
| S1205R | S. cerevisiae | • S1202R: Reduced adenylate kinase | Bhaskara et al. (2007) | |
| S793R | P. furiosus | • S793R: Deficient in ATP-dependent dimer formation and ATP binding | ||
| S1202R | H. sapiens | • S1202R and S1205R: Low level of adenylate kinase | ||
| • S1205R: Telomere shortening, not support spore viability | ||||
| Signature motif and Q loop | S793R, Q140H | P. furiosus | • S793R: Analogs to the mutation in CFTR (S549R) gene that results cystic fibrosis | Moncalian et al. (2004) |
| • S793R: Prevented ATP binding | ||||
| S1205R | S. cerevisiae | • S1205R S. cerevisiae: Failed to complement Rad50 deletion strain in DNA repair assay | ||
| • S783R and Q140H: Halted ATP-dependent activities | ||||
| ATPase domain | R1093 (stop) c.3939A/T | H. sapiens | • Nijmegen breakage syndrome like disorder (NBSLD) | Waltes et al. (2009) |
Note:
HR, homologous recombination; NHEJ, non-homologous end joining repair; CFTR, cystic fibrosis transmembrane conductance regulator; ATP, adenosine tri-phosphate; ATM, ataxia-telangiectasia mutated; ATR, ATM-and Rad3-Related. Refer to Table S2 for the description of all mutations.