Experimental limits of PCR analysis of (CA)n repeat alterations

Foucault, Frédéric; Praz, Françoise; Jaulin, Christian; Amor‐Guéret, Mounira

doi:10.1016/0168-9525(96)99996-5

Cited by 31 publications

(26 citation statements)

References 9 publications

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“…Available evidence indicates that mutations of the cysteine residues in the zinc finger motif in most RecQ family of helicases can have dramatic effects on the enzymatic activities. Two disease-causing Bloom missense mutations map to Cys 1036 and Cys 1055 , respectively (32,33), among four conserved cysteine residues in the zinc finger motif. In vitro analysis shows that these mutations abolish BLM ATPase and helicase activities (34).…”

Section: Discussionmentioning

confidence: 99%

The Zinc Finger Motif of Escherichia coli RecQ Is Implicated in Both DNA Binding and Protein Folding

Liu¹,

Rigolet²,

Dou³

et al. 2004

Journal of Biological Chemistry

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The RecQ family of DNA helicases has been shown to be important for the maintenance of genomic integrity. Mutations in human RecQ genes lead to genomic instability and cancer. Several RecQ family of helicases contain a putative zinc finger motif of the C 4 type at the C terminus that has been identified in the crystalline structure of RecQ helicase from Escherichia coli. To better understand the role of this motif in helicase from E. coli, we constructed a series of single mutations altering the conserved cysteines as well as other highly conserved residues. All of the resulting mutant proteins exhibited a high level of susceptibility to degradation, making functional analysis impossible. In contrast, a double mutant protein in which both cysteine residues Cys 397 and Cys 400 in the zinc finger motif were replaced by asparagine residues was purified to homogeneity. Slight local conformational changes were detected, but the rest of the mutant protein has a well defined tertiary structure. Furthermore, the mutant enzyme displayed ATP binding affinity similar to the wild-type enzyme but was severely impaired in DNA binding and in subsequent ATPase and helicase activities. These results revealed that the zinc finger binding motif is involved in maintaining the integrity of the whole protein as well as DNA binding. We also showed that the zinc atom is not essential to enzymatic activity.The transient formation of single-stranded DNA (ssDNA) 1 intermediate is essential to all aspects of DNA metabolism including DNA replication, recombination, and repair. The unwinding and separation of the individual strands of doublestranded DNA (dsDNA) is catalyzed by a class of specialized enzymes known as DNA helicases (1, 2). These enzymes function as molecular motors that use the energy released from the hydrolysis of ATP to unwind and translocate along DNA in a sequential fashion (3-5). These ubiquitous enzymes have been identified in all living organisms from virus to human. It appears that they evolved from a common ancestor (6).The RecQ helicase family is critical to the maintenance of genomic stability in prokaryotes and eukaryotes (7). Mutations of RecQ genes can lead to genomic instability and several human diseases including the Bloom and Werner syndromes (8). Recently, it has been shown that the tumor suppressor BRCA1-associated protein, BACH1, which shares homologies with other members of the RecQ family, possesses ATPase and helicase activities (9). The mutant BACH1 participates directly in breast and ovarian cancer development (9). The RecQ helicase from Escherichia coli is the prototype helicase of this family (10) and has been shown to initiate homologous recombination as well as suppress illegitimate recombination (11, 12).The RecQ helicase family members contain a helicase domain characterized by the presence of seven so-called "helicase" motifs necessary for using energy derived from ATP binding and hydrolysis to unwind DNA (13,14). Sequence analyses revealed that all of the RecQ helicases contain a C-terminal e...

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Section: Discussionmentioning

confidence: 99%

The Zinc Finger Motif of Escherichia coli RecQ Is Implicated in Both DNA Binding and Protein Folding

Liu¹,

Rigolet²,

Dou³

et al. 2004

Journal of Biological Chemistry

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“…86 A significant drawback of WGA techniques is that amplification of repetitive DNA sequences, such as short tandem repeats, is error prone if performed on WGA products. 86,97 In some studies over 50% of fragments amplified differed from their expected size presumably due to the uniformly low temperatures needed for WGA which could allow slippage of the DNA chain during product generation. 86 Such errors would currently rule out the use of WGA for the clinical diagnosis of triplet repeat expansion diseases or diagnoses based on linkage analysis with STRs.…”

Section: Whole Genome Amplificationmentioning

confidence: 99%

Molecular Diagnostics in Preimplantation Genetic Diagnosis

Thornhill

Snow

2002

The Journal of Molecular Diagnostics

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“…In many cases this required 10^100 replicates for a certain individual at a certain locus. Alleles that were seen only once were considered to be artefactual (Foucault et al 1996) and were ignored. Ampli¢cations with a heterozygous score for a given locus were veri¢ed in at least three independent PCRs, in most cases from independent probes of the individual.…”

Section: (E) Microsatellite Analysismentioning

confidence: 99%

Intracommunity relationships, dispersal pattern and paternity success in a wild living community of Bonobos (Pan paniscus) determined from DNA analysis of faecal samples

Gerloff¹,

Hartung

Fruth

et al. 1999

Proc. R. Soc. Lond. B

252

162

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Di¡erences in social relationships among community members are often explained by di¡erences in genetic relationships. The current techniques of DNA analysis allow explicit testing of such a hypothesis. Here, we have analysed the genetic relationships for a community of wild bonobos (Pan paniscus) using nuclear and mitochondrial DNA markers extracted from faecal samples. Bonobos show an opportunistic and promiscuous mating behaviour, even with mates from outside the community. Nonetheless, we ¢nd that most infants were sired by resident males and that two dominant males together attained the highest paternity success. Intriguingly, the latter males are the sons of high-ranking females, suggesting an important in£uence of mothers on the paternity success of their sons. The molecular data support previous inferences on female dispersal and male philopatry. We ¢nd a total of ¢ve di¡erent mitochondrial haplotypes among 15 adult females, suggesting a frequent migration of females. Moreover, for most adult and subadult males in the group we ¢nd a matching mother, while this is not the case for most females, indicating that these leave the community during adolescence. Our study demonstrates that faecal samples can be a useful source for the determination of kinship in a whole community.

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Experimental limits of PCR analysis of (CA)n repeat alterations

Cited by 31 publications

References 9 publications

The Zinc Finger Motif of Escherichia coli RecQ Is Implicated in Both DNA Binding and Protein Folding

The Zinc Finger Motif of Escherichia coli RecQ Is Implicated in Both DNA Binding and Protein Folding

Molecular Diagnostics in Preimplantation Genetic Diagnosis

Intracommunity relationships, dispersal pattern and paternity success in a wild living community of Bonobos (Pan paniscus) determined from DNA analysis of faecal samples

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