Alu-mediated nonallelic homologous and nonhomologous recombination in the BMPR2 gene in heritable pulmonary arterial hypertension

Kataoka, Masaharu; Aimi, Yuki; Yanagisawa, Ryoji; Ono, Masae; Oka, Akira; Fukuda, Keiichi; Yoshino, Hideaki; Satoh, Tsuyoshi; Gamou, Shinobu

doi:10.1038/gim.2013.41

Cited by 25 publications

(22 citation statements)

References 17 publications

(15 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this study, we have shown in both RI assay and chromosomal DSB joining assay that SSA-type recombination relies on ≥13 bp homology, which contrasts with the 2–6 bp preference seen in A-EJ. Our findings described here shed light on the mechanism of SSA (between Alu elements), which, albeit normally suppressed, has been reported to cause deleterious outcomes involving deletions and translocations545556.…”

Section: Discussionmentioning

confidence: 57%

Dual loss of human POLQ and LIG4 abolishes random integration

2017

View full text Add to dashboard Cite

Homologous recombination-mediated gene targeting has greatly contributed to genetic analysis in a wide range of species, but is highly inefficient in human cells because of overwhelmingly frequent random integration events, whose molecular mechanism remains elusive. Here we show that DNA polymerase θ, despite its minor role in chromosomal DNA repair, substantially contributes to random integration, and that cells lacking both DNA polymerase θ and DNA ligase IV, which is essential for non-homologous end joining (NHEJ), exhibit 100% efficiency of spontaneous gene targeting by virtue of undetectable levels of random integration. Thus, DNA polymerase θ-mediated end joining is the sole homology-independent repair route in the absence of NHEJ and, intriguingly, their combined absence reveals rare Alu-Alu recombination events utilizing a stretch of homology. Our findings provide new insights into the mechanics of foreign DNA integration and the role of DNA polymerase θ in human genome maintenance.

show abstract

Section: Discussionmentioning

confidence: 57%

Dual loss of human POLQ and LIG4 abolishes random integration

2017

View full text Add to dashboard Cite

show abstract

“…16 In that study, we found that BMPR2 point mutations and exon deletions may account for at least some of the BMPR2 mutations associated with PAH in a Japanese population, and that the exon deletions arise not only via Alumediated non-allelic homologous recombination, but also via nonhomologous recombination. 16 Herein we summarize updated genetic analysis of BMPR2 in Japanese PAH patients and their families. These patients are from single ethnic group and mostly reside in metropolitan Tokyo and the surrounding area, thus sharing many common factors and resulting in a racially, ethnically, and socially homogeneous population.…”

Section: Bmpr2 In Pulmonary Arterial Hypertensionmentioning

confidence: 89%

“…It is interesting to explore and review the distribution of reported 16 Machado et al 26,27 16 Machado et al, 26 Kabata et al 28 11 Machado et al, 17,26,27 Momose et al, 25 Elliott et al, 29 Sztrymf et al, 30 Wang et al, 31 Girerd et al, 32 Liu et al, 33 Pfarr et al 34 16 Machado et al, 26 Kabata et al 28 16 Machado et al 26,27 (rs863223424) Nonsense 7 c.961C>T p.Arg321* HPAH F Machado et al, 26,27 Wang et al, 31 Girerd et al, 32 Pfarr et al, 34 Koehler et al 35 29 Girerd et al, 32 Liu et al, 33 Thomson et al, 36 Sankelo et al, 37 Portillo et al 38 16 Machado et al, 26 Kabata et al 28 9 Machado et al, 26,27,39 Liu et al, 33 Pfarr et al, 34 Sankelo et al, 37 Portillo et al, 38 16 Machado et al, …”

Section: Functional Analysis For Pathogenicity Using Common Databasesmentioning

confidence: 99%

Genetics in pulmonary arterial hypertension in a large homogeneous Japanese population

et al. 2018

View full text Add to dashboard Cite

Pulmonary arterial hypertension (PAH) is a rare but serious disease with a grave prognosis. Bone morphogenetic protein type 2 receptor (BMPR2) gene is a strong pathogenic factor for PAH. As a collaborative team from Kyorin University and Keio University in Japan, we have analyzed the BMPR2 gene in 356 probands and more than 50 family members, including secondary patients. Importantly, the study population is a racially, ethnically, and socially homogeneous population. In PAH patients, there is a high incidence of unique mutations in BMPR2, and several mutations are frequently observed in the Japanese population, suggesting that these common and recurring mutations may be highly pathogenic or have high penetrance, explaining why they are found frequently throughout the world. We have also mapped each breakpoint of exonic deletions/duplications and found that most break and rejoining points are in the Alu elements. Reviewing the distribution of the reported mutations on each exon of BMPR2 revealed that the number and frequency of mutations are imbalanced among exons. The penetrance of BMPR2 gene mutations was 3-fold higher in females than males. Full elucidation of BMPR2-mediated pathogenic mechanisms in PAH requires persistent efforts to achieve precision or individualized medicine as a therapeutic strategy for PAH.

show abstract

“…Another intriguing possibility is that in the absence of Rad54 paralogs, HR‐independent gene targeting could somehow take place, for example, in a manner depending on limited sequence homologies or via homeologous sequences . As such, Alu–Alu recombination has been reported to occur in human cells, causing deleterious outcomes such as deletions, translocations, or copy number variations . In the light of gene targeting, because of the abundance of Alu or other repetitive DNA sequences in the human genome , targeting vectors typically contain large amounts of repetitive DNA in their homology arms .…”

Section: Discussionmentioning

confidence: 99%

Mechanistic basis for increased human gene targeting by promoterless vectors—roles of homology arms and Rad54 paralogs

2017

View full text Add to dashboard Cite

Gene targeting by homologous recombination provides the definitive tool for analyzing gene function. Promoterless vectors, which do not possess a promoter to drive marker gene expression, confer higher targeting efficiencies than conventional vectors due to the reduced number of drug-resistant clones. We here show that gene-targeting efficiency is typically ≥ 25% with the use of exon-trapping-type promoterless vectors in a human diploid cell line, Nalm-6. The efficiency of exon-trapping gene targeting was correlated with the level of target gene expression when a 2A peptide sequence was linked to the marker gene. Intriguingly, total arm length was not necessarily a determinant of targeting efficiency, as longer arms tend to enhance both homologous (targeted) and nonhomologous (nontargeted) integration of the vector; rather, the presence of an exon in the 5' arm led to a decreased targeting efficiency. Strikingly, loss of Rad54 did not severely affect the targeting efficiency of exon-trap vectors. Moreover, additional deletion of the Rad54 paralog Rad54B had limited impact on the high-efficiency gene targeting. These results indicate that targeted integration occurs in human cells even when both Rad54 and Rad54B are missing. These studies provide additional important insight into the contribution of various DNA repair factors on the targeting mechanics.

show abstract

Alu-mediated nonallelic homologous and nonhomologous recombination in the BMPR2 gene in heritable pulmonary arterial hypertension

Cited by 25 publications

References 17 publications

Dual loss of human POLQ and LIG4 abolishes random integration

Dual loss of human POLQ and LIG4 abolishes random integration

Genetics in pulmonary arterial hypertension in a large homogeneous Japanese population

Mechanistic basis for increased human gene targeting by promoterless vectors—roles of homology arms and Rad54 paralogs

Contact Info

Product

Resources

About