Advances in Mutagenesis Research

205 Pages·1991·7.955 MB·English

by  M. Frankenberg-Schwager, D. Frankenberg (auth.), Professor Dr. Günter Obe (eds.)

3
Advances
in Mutagenesis Research _____
Editor-in-Chief
O. Obe, Essen
Editorial Board
H. J. Evans, Edinburgh
A. T. Natarajan, Leiden
H. S. Rosenkranz, Cleveland
F. H. Sobels, Leiden
T. Sugimura, Tokyo
Advances
in Mutagenesis
Research 3
Edited by O. Obe
With Contributions by
H. H. Evans D. Frankenberg
M. Frankenberg-Schwager E. Gebhart
W. Kohnlein R. H. Nussbaum
D. G. Papworth J. R. K. Savage
R.-D. Wegner
With 58 Figures
Springer-Verlag
Berlin Heidelberg New York
London Paris Tokyo
Hong Kong Barcelona
Budapest
Professor Dr. GONTER 0BE
FB9 der Universitat
Gesamthochschule Essen
UniversitatsstraBe 5
Postfach 103764
4300 Essen 1, FRG
ISBN-13: 978-3-642-76234-5 e-ISBN-13: 978-3-642-76232-1
DOl: 10.1007/978-3-642-76232-1
Library of Congress Catalog Card Number 89-640326
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Foreword to the Series
Mutations are permanent changes in the genetic material. These
changes can comprise single genes (gene mutations), the structure
of the chromosomes (chromosome mutations), or the number of
the chromosomes (genome mutations). Since H. J. Muller present
ed his paper The problem oj genic modification at the 5th Interna
tional Congress of Genetics in Berlin on the 15th of September,
1927, in which he brilliantly showed that X-rays induce mutations
in the fruit fly Drosophila, we have learnt that a plethora of agents,
including ionizing and nonionizing radiations, chemicals, and
viruses, can induce mutations. In most of the cases, induced muta
tions are deleterious to the cells or the organisms in which they oc
cur, and we cannot justify damaging the genetic material of organ
isms, including ourselves, by introducing man-made mutagenic
agents into the environment. To prevent this, chemicals must be
tested for their possible mutagenicity in a variety of test systems
before they can be used. This has opened a field of applied genetic
research, namely, genetic toxicology. Comparative analyses led to
the concept that mutagenic agents can be expected to be also car
cinogenic. The theory of the origin of cancer by mutations has
gained experimental proof by the finding that oncogenes, when
changed by mutations, can give rise to cancer.
Basic research in the field of mutation research has unraveled
some of the molecular mechanisms underlying the origin of muta
tions and the complex reaction of cells to induced changes in their
DNA. These cellular reactions can eventually lead to the restora
tion of the original structure of the DNA, but, via misrepair, can
also give rise to mutations.
There are still many open questions. The molecular mecha
nisms leading to mutations are only partially known. In view of
the fact that about 6 in 1000 newborn children have a chromo
somal alteration, it would be especially important to understand
how chromosome and genome mutations are produced.
Molecular changes in the DNA and the reaction of the cells to
such changes result in typical mutation rates which reflect the
evolutionary history of the organisms in question. Mutations are
one of the sources of variability which is the prerequisite for
natural selection and for evolution; but since mutations can also
VI Foreword to the Series
result in various deleterious effects, such as hereditary diseases, a
population can only survive when the mutation rates are not too
high and not too low, i.e., mutation rates are delicately balanced.
Elevations of the mutation rates would have considerable conse
quences. It would lead to an increase in the frequencies of cancers
and would represent a great risk for the evolutionary future of a
species; a scenario in which humans are fully included.
In view of these implications, mutation research has two aims:
1. To understand the molecular mechanisms leading to mutations
and
2. to prevent a thoughtless introduction of mutagenic agents into
our environment.
Both aspects, namely basic and applied ones, will be treated in the
series Advances in Mutagenesis Research. The articles will deal
with current developments in the field of mutation research and
will help the reader to orient himself in this centrally important
area of biology.
Prof. Dr. GONTER aBE
Contents
Rejoining of Radiation-Induced
DNA Double-Strand Breaks in Yeast
M. FRANKENBERG-SCHWAGER and D. FRANKENBERG
(With 19 Figures) ................................... 1
1 Introduction .................................... .
2 Measurement of DNA Double-Strand Breaks
by the Neutral Sucrose Sedimentation Method ....... 3
3 Induction of DNA Double-Strand Breaks in Yeast
Irradiated with Ionizing Radiations:
Effect of Linear Energy Transfer and Oxia . . . . . . . . . 5
4 DNA Double-Strand Breaks as Critical Lesions
for Yeast Cell Killing ............................. 5
5 Rejoining of DNA Double-Strand Breaks
Induced in Yeast by Sparsely Ionizing Radiation ..... 9
6 Rejoining of DNA Double-Strand Breaks
Induced in Yeast by 3.5 MeV a-Particles ............. 16
7 DNA-Double-Strand Breaks Induced in Anoxic Yeast
Show Different Rejoining Kinetics Compared to
Oxically Induced Double-Strand Breaks ............. 20
8 Concluding Remarks ...... . . . . . . . . . . . . . . . . . . . . . . . . 24
References ......................................... 25
Cellular and Molecular Effects of Radon
and Other Alpha Particle Emitters
H.H. EVANS (With 10 Figures) ....................... 28
1 Inhalation of Radon Induces Lung Cancer .......... 28
2 Energy Deposition by Alpha Particles ............... 29
3 Evidence That Irreparable Lesions Are Induced
by Alpha Particles ................................ 29
4 Induction of DNA Double-Strand Breaks
by Alpha Radiation .............................. 33
5 Chromosomal Damage Induced by Alpha Radiation .. 35
6 Mutagenicity of Alpha Radiation ................... 38
7 Characterization of Mutational Lesions
Induced by Alpha Radiation ....................... 40
VIII Contents
8 Oncogenic Transformation Induced by
Alpha Radiation ................................. 42
9 Role of Ionizing Radiation in Oncogenic Transformation 43
References ......................................... 46
Reassessment of Radiogenic Cancer Risk and Mutagenesis
at Low Doses of Ionizing Radiation
W. KOHNLEIN and R.H. NUSSBAUM (With 7 Figures) ... 53
1 Introduction ..................................... 53
2 Low Dose Mutagenicity Studies .................... 54
3 The Present Basis of Radiation Protection Guidelines. 62
4 Epidemiological Studies Limited to Low Doses ....... 63
5 The A-Bomb Survivor Study......... .............. 64
6 Results .......................................... 73
7 Discussion ....................................... 75
8 Comparison with Other Low-Dose Epidemiological
Studies.......................................... 76
9 Conclusion ...................................... 77
Appendix: Lifetime Cancer Risk Projection ............ 78
References ......................................... 78
Chromosomal Instability Syndromes in Man
R.-D. WEGNER (With 15 Figures) ..................... 81
1 Introduction ..................................... 81
2 Clinical Characteristics ............................ 84
3 Cytogenetic Characteristics ........................ 88
4 Heterozygote Detection and Prenatal Diagnosis ...... 107
5 Genetic Heterogeneity and Molecular Studies ........ 111
6 Conclusions ..................................... 117
References ......................................... 117
Chromosomal Changes in Nonneoplastic Somatic Cells
of Cancer Patients:
Indication of a Predisposing Chromosomal Instability?
E. GEBHART (With 2 Figures) ........................ 131
1 Introduction ..................................... 131
2 Methodological Approaches ....................... 133
3 Results .......................................... 135
4 Conclusions ..................................... 151
References ......................................... 153
Contents IX
Excogitations About the Quantification
of Structural Chromosomal Aberrations
J.R.K. SAVAGE and D.G. PAPWORTH (With 5 Figures) 162
1 Introduction ..................................... 162
2 Necessary Prerequisites ............................ 163
3 The Ideal Situation ............................... 166
4 Imprecision: The Effect of Cell Kinetics ............. 173
5 The Influence of Mitotic Delay .................... 177
6 Mitigating Problems by Cohort Analysis ............ 180
7 Concluding Comments............................ 183
References ......................................... 184
Subject Index ...................................... 190
Rejoining of Radiation-Induced DNA
Double-Strand Breaks in Yeast
M. FRANKENBERG-SCHWAGER and D. FRANKENBERG
Contents
1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 1
2 Measurement of DNA Double-Strand Breaks by the Neutral Sucrose Sedimentation Method 3
3 Induction of DNA Double-Strand Breaks in Yeast Irradiated with Ionizing Radiations:
Effect of Linear Energy Transfer and Oxia . . . . . . . . . . . . . . . . . . . . . . . . . . .. 5
4 DNA Double-Strand Breaks as Critical Lesions for Yeast Cell Killing. . . . . . . . . . . .. 5
5 Rejoining of DNA Double-Strand Breaks Induced in Yeast by Sparsely Ionizing Radiation 9
5.1 Double-Strand Break Rejoining Requires Homologous DNA Sequences. . 9
5.2 Double-Strand Break Rejoining Occurs in Growth Medium ................... 10
5.3 Double-Strand Break Rejoining Also Occurs in Nongrowth Medium .............. 10
5.4 The Linear Relationship Between Induced Double-Strand Breaks and Dose Is Converted
into a Dose-Squared Relationship Between Unrejoined Double-Strand Breaks and Dose 11
5.5 The Kinetics of Double-Strand Break Rejoining Is Biphasic and Unsaturated . . . . 12
5.6 Double-Strand Breaks Are Rejoined During Irradiation at Low Dose Rate. . . . . . . 14
6 Rejoining of DNA Double-Strand Breaks Induced in Yeast by 3.5 MeV a-Particles . 16
6.1 Split Dose Rejoining of a-Particle Induced Double-Strand Breaks . . . . . . . . . . 16
6.2 Unrejoined Double-Strand Breaks Induced by a-Particles Show a Dose-Squared Term. 16
6.3 Double-Strand Break Rejoining Determines the Relative Biological Efficiency
of a-Particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
6.4 The Kinetics of Rejoining of a-Particle Induced Double-Strand Breaks Is Monophasic
and Unsaturated ....................................... . 18
7 DNA Double-Strand Breaks Induced in Anoxic Yeast Show Different Rejoining Kinetics
Compared to Oxically Induced Double-Strand Breaks . . . . . . . . . 20
7.1 Formation of Secondary Double-Strand Breaks During Incubation
of Anoxically Irradiated Yeast Cells in Nongrowth Medium . . . . . 21
7.2 Rejoining Kinetics of Anoxically Induced Double-Strand Breaks. . . 22
7.3 The Oxygen Enhancement Ratio for Induced Double-Strand Breaks Is Higher Than That
for Unrejoined Double-Strand Breaks. . 23
8 Concluding Remarks . . 24
References .................. . 25
1 Introduction
Among the various lesions detected in the DNA of irradiated cells, DNA double
strand breaks (DSB) and bulky lesions (denatured regions comprising at least three
unpaired bases) lead to structural disturbances which may have importa'nt biolog
ical consequences (Fig. 1). While little is known about radiation-induced bulky
lesions in eukaryotes (Geigl 1987), intensive investigations on radiation-induced
Gesellschaft fUr Strahlen-und Umweltforschung mbH. Institut fUr Biophysikalische Strahlenforschung.
Paul-Ehrlich -Str. 20, 6000 Frankfurt 70, FRG
2 M. Frankenberg-Schwager and D. Frankenberg
- -base alteration
crosslink
, __ base
detachment
bulky
lesion
Fig. 1. Schematic illustralion of lesions
induced in cellular DNA by ionizing ra
diation (Frankenberg-Schwager 1989)
DNA DSB have been performed in recent years. The unicellular yeast
Saccharomyces cerevisiae has contributed significantly of the understanding of
the role of DSB in irradiated eukaryotic cells. This organism proved to be
especially suitable to study induction and rejoining ofDSB at biologically relevant
radiation doses because of the small size of its chromosomal DNA molecules,
allowing the detection of few DSB per cell by the neutral sucrose sedimentation
technique. A haploid yeast cell contains per nucleus 16 chromosomes of varying
DNA length ranging from 150 to about 2500 kb pairs. The yeast chromatin is
organized in nucleosomes containing the four core histones H2A, H2B, H3, and
H4, but clear evidence for the HI histone is missing (see Fangman and Zakian
1981). Unlike mammalian cells, radiation-sensitive yeast mutants exist which do
not show a detectable rejoining of radiation-induced DSB. These mutants are
valuable tools to investigate the biological relevance of DSB. Especially helpfull
in this respect are mutants which are temperature conditional for DSB rejoining.
Although yeast is an ideal organism to study the fate of DNA DSB and their
role in cell killing at biologically relevant doses, cytological investigations are
difficult to perform because of the small size of the yeast chromosomes. However,
in mammalian cells evidence exists that DNA DSB produced by restriction
endonucleases give rise to chromosome aberrations which are of the same type as
those induced by X-rays (Bryant 1984; Natarajan and Obe 1984). Experimental
data are accumulating that radiation-induced DSB may lead to chromosome