The induction of specific-locus mutations with N-propyl-N-nitrosourea in stem-cell spermatogonia of mice

Murota, Tetsuro; Shibuya, Tohru

doi:10.1016/0165-7992(91)90085-i

Cited by 10 publications

(3 citation statements)

References 10 publications

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“…As anticipated, mutations in the Pig‐a gene were identified in bulk‐sorted CD59‐deficient BMEs from rats treated with both mutagens. PCZ and PNU are known as powerful mutagens in vitro and in vivo, and as rodent carcinogens (Lee and Dixon, 1978; Nishi et al ., 1984; Suter, 1987; Ogiu et al ., 1988; Murota and Shibuya, 1991; Myhr, 1991; Suzuki et al ., 1999a; 1999b; Phonethepswath et al ., 2013; Chung et al ., 2018). Surprisingly, very little information is available on the mutational spectra induced by PCZ and PNU.…”

Section: Discussionmentioning

confidence: 99%

CD59‐deficient bone marrow erythroid cells from rats treated with procarbazine and propyl‐nitrosourea have mutations in thePig‐agene

Revollo

Dad

Pearce

et al. 2020

Environ and Mol Mutagen

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Procarbazine (PCZ) and N-propyl-N-nitrosourea (PNU) are rodent mutagens and carcinogens. Both induce GPI-anchored marker-deficient mutant-phenotype red blood cells (RBCs) in the flow cytometry-based rat RBC Pig-a assay. In the present study, we traced the origin of the RBC mutant phenotype by analyzing Pig-a mutations in the precursors of RBCs, bone marrow erythroid cells (BMEs). Rats were exposed to a total of 450 mg/kg PCZ hydrochloride or 300 mg/kg PNU, and bone marrow was collected 2, 7, and 10 weeks later. Using a flow cell sorter, we isolated CD59-deficient mutant-phenotype BMEs from PCZ-and PNU-treated rats and examined their endogenous X-linked Pig-a gene by next generation sequencing. Pig-a mutations consistent with the properties of PCZ and PNU were found in sorted mutant-phenotype BMEs. PCZ induced mainly A > T transversions with the mutated A on the nontranscribed strand of the Pig-a gene, while PNU induced mainly T > A transversions with the mutated T on the nontranscribed strand. The treatmentinduced mutations were distributed across the protein coding sequence of the Pig-a gene. The causal relationship between BMEs and RBCs and the agent-specific mutational spectra in CD59-deicient BMEs indicate that the rat RBC Pig-a assay, scoring CD59-deficient mutant-phenotype RBCs in peripheral blood, detects Pig-a gene mutation. K E Y W O R D S flow cytometry, glycosyl phosphatidylinositol, red blood cells, sequencing 1 | INTRODUCTION Mutation in the mammalian endogenous X-linked Pig-a gene can disrupt glycosyl phosphatidylinositol (GPI) synthesis, resulting in a mutant phenotype that is characterized by a deficiency in specific GPI-anchored surface markers (Kinoshita et al., 1997; Kinoshita, 2014). Since mutation in the human PIG-A gene is associated with a disease, paroxysmal nocturnal hemoglobinuria, diagnostic assays have been designed for detecting mutant-phenotype cells

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Section: Discussionmentioning

confidence: 99%

CD59‐deficient bone marrow erythroid cells from rats treated with procarbazine and propyl‐nitrosourea have mutations in thePig‐agene

Revollo

Dad

Pearce

et al. 2020

Environ and Mol Mutagen

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“…Here, we have improved the workflow for sequencing the DNA of T‐cell clones as an approach for assessing whole genome mutational load; and we have derived mutational signatures for a reference mutagen, N ‐propyl‐ N ‐nitrosourea (PNU), and a genotoxic antineoplastic agent, procarbazine (PCZ). Both compounds are known as powerful mutagens (Chung et al, 2018; Clive et al, 1988; Holme et al, 1989; Hoorn et al, 1993; Lee & Dixon, 1978; Murota & Shibuya, 1991; Myhr, 1991; Ogiu et al, 1988; Suzuki, Itoh, et al, 1999; Suzuki, Uno, et al, 1999), yet their mutational properties are not well characterized. Our data can serve as a starting point for further refinement of sequencing strategies and improvement of the sensitivity and specificity of the whole genome mutation detection approach.…”

Section: Introductionmentioning

confidence: 99%

Mutational signatures in T‐lymphocytes of rats treated with N‐propyl‐N‐nitrosourea and procarbazine

Revollo

McKinzie

Robison

et al. 2021

Environ and Mol Mutagen

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We have used whole genome sequencing (WGS) to determine mutational signatures induced in the T‐cells of rats treated in vivo with N‐propyl‐N‐nitrosourea (PNU) or procarbazine (PCZ). The signatures from the treated rats were different from the signature of background mutations. The main component of the spontaneous T‐cell mutational signature was C➔T transition with all other single base substitutions evenly distributed. The PNU‐induced mutational signature showed relatively equal contributions from C➔T and T➔C transitions, and T➔A transversions. The PCZ‐induced signature was characterized by T➔C transitions, T➔A and, to a smaller extent, T➔G transversions. C➔G transversions were infrequent in either the PNU or PCZ signatures. WGS not only allowed mutational signature detection, but also measured quantitative responses to mutagen treatment: 10–40× increases in the number of mutations per clone were detected in T‐cell clones from treated rats. The overall strand specificity of induced mutations for annotated rat genes was comparable to the strand specificity of mutations determined previously for the endogenous X‐linked Pig‐a gene. Our results provide valuable reference data for future applications of WGS in safety research and risk assessment.

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“…1981 Murota et al, 1982;Murota and Shibuya, 1983;1991;Shibuya et al, 1993;1996;Murota, 199519721972198919801985Mutation Research 1977MMS 19881960FDA in vivo 19701977 54…”

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confidence: 99%

Achievements of the late Kiyoshi Tutikawa, a mouse geneticist

Kikuchi¹

2004

Environ. Mutagen Res.

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The induction of specific-locus mutations with N-propyl-N-nitrosourea in stem-cell spermatogonia of mice

Cited by 10 publications

References 10 publications

CD59‐deficient bone marrow erythroid cells from rats treated with procarbazine and propyl‐nitrosourea have mutations in thePig‐agene

CD59‐deficient bone marrow erythroid cells from rats treated with procarbazine and propyl‐nitrosourea have mutations in thePig‐agene

Mutational signatures in T‐lymphocytes of rats treated with N‐propyl‐N‐nitrosourea and procarbazine

Achievements of the late Kiyoshi Tutikawa, a mouse geneticist

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