Impaired DNA replication prompts deletions within palindromic sequences, but does not induce translocations in human cells

Hiroki Kurahashi, Hidehito Inagaki, Takema Kato, Eriko Hosoba, Hiroshi Kogo, Tamae Oe, Makiko Tsutsumi, Hasbaira Bolor, Maoqing Tong, Beverly S. Emanuel

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Palindromic regions are unstable and susceptible to deletion in prokaryotes and eukaryotes possibly due to stalled or slow replication. In the human genome, they also appear to become partially or completely deleted, while two palindromic AT-rich repeats (PATRR) contribute to known recurrent constitutional translocations. To explore the mechanism that causes the development of palindrome instabilities in humans, we compared the incidence of de novo translocations and deletions at PATRRs in human cells. Using a highly sensitive PCR assay that can detect single molecules, de novo deletions were detected neither in human somatic cells nor in sperm. However, deletions were detected at low frequency in cultured cell lines. Inhibition of DNA replication by administration of siRNA against the DNA polymerase alpha 1 (POLA1) gene or introduction of POLA inhibitors increased the frequency. This is in contrast to PATRR-mediated translocations that were never detected in similar conditions but were observed frequently in human sperm samples. Further deletions were found to take place during both leading- and lagging-strand synthesis. Our data suggest that stalled or slow replication induces deletions within PATRRs, but that other mechanisms might contribute to PATRR-mediated recurrent translocations in humans.

Original languageEnglish
Pages (from-to)3397-3406
Number of pages10
JournalHuman molecular genetics
Volume18
Issue number18
DOIs
Publication statusPublished - 04-09-2009

Fingerprint

DNA Replication
Spermatozoa
DNA Polymerase I
Human Genome
Eukaryota
Small Interfering RNA
Cultured Cells
Cell Line
Polymerase Chain Reaction
Incidence
Genes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Cite this

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title = "Impaired DNA replication prompts deletions within palindromic sequences, but does not induce translocations in human cells",
abstract = "Palindromic regions are unstable and susceptible to deletion in prokaryotes and eukaryotes possibly due to stalled or slow replication. In the human genome, they also appear to become partially or completely deleted, while two palindromic AT-rich repeats (PATRR) contribute to known recurrent constitutional translocations. To explore the mechanism that causes the development of palindrome instabilities in humans, we compared the incidence of de novo translocations and deletions at PATRRs in human cells. Using a highly sensitive PCR assay that can detect single molecules, de novo deletions were detected neither in human somatic cells nor in sperm. However, deletions were detected at low frequency in cultured cell lines. Inhibition of DNA replication by administration of siRNA against the DNA polymerase alpha 1 (POLA1) gene or introduction of POLA inhibitors increased the frequency. This is in contrast to PATRR-mediated translocations that were never detected in similar conditions but were observed frequently in human sperm samples. Further deletions were found to take place during both leading- and lagging-strand synthesis. Our data suggest that stalled or slow replication induces deletions within PATRRs, but that other mechanisms might contribute to PATRR-mediated recurrent translocations in humans.",
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Impaired DNA replication prompts deletions within palindromic sequences, but does not induce translocations in human cells. / Kurahashi, Hiroki; Inagaki, Hidehito; Kato, Takema; Hosoba, Eriko; Kogo, Hiroshi; Oe, Tamae; Tsutsumi, Makiko; Bolor, Hasbaira; Tong, Maoqing; Emanuel, Beverly S.

In: Human molecular genetics, Vol. 18, No. 18, 04.09.2009, p. 3397-3406.

Research output: Contribution to journalArticle

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T1 - Impaired DNA replication prompts deletions within palindromic sequences, but does not induce translocations in human cells

AU - Kurahashi, Hiroki

AU - Inagaki, Hidehito

AU - Kato, Takema

AU - Hosoba, Eriko

AU - Kogo, Hiroshi

AU - Oe, Tamae

AU - Tsutsumi, Makiko

AU - Bolor, Hasbaira

AU - Tong, Maoqing

AU - Emanuel, Beverly S.

PY - 2009/9/4

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N2 - Palindromic regions are unstable and susceptible to deletion in prokaryotes and eukaryotes possibly due to stalled or slow replication. In the human genome, they also appear to become partially or completely deleted, while two palindromic AT-rich repeats (PATRR) contribute to known recurrent constitutional translocations. To explore the mechanism that causes the development of palindrome instabilities in humans, we compared the incidence of de novo translocations and deletions at PATRRs in human cells. Using a highly sensitive PCR assay that can detect single molecules, de novo deletions were detected neither in human somatic cells nor in sperm. However, deletions were detected at low frequency in cultured cell lines. Inhibition of DNA replication by administration of siRNA against the DNA polymerase alpha 1 (POLA1) gene or introduction of POLA inhibitors increased the frequency. This is in contrast to PATRR-mediated translocations that were never detected in similar conditions but were observed frequently in human sperm samples. Further deletions were found to take place during both leading- and lagging-strand synthesis. Our data suggest that stalled or slow replication induces deletions within PATRRs, but that other mechanisms might contribute to PATRR-mediated recurrent translocations in humans.

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