Table Of ContentThis is a volume in
CELL BIOLOGY
A series of monographs
Editors: D. E. Buetow, I. L. Cameron, G. M. Padilla, and A. M. Zimmerman
A complete list of the books in this series appears at the end of the volume.
PREMATURE
CHROMOSOME
CONDENSATION
Application in Basic,
Clinical, and
Mutation Research
Edited by
Potu Ν. Rao
Department of Developmental Therapeutics
The University of Texas System Cancer Center
M. D. Anderson Hospital and Tumor Institute
Houston, Texas
Robert T. Johnson
Department of Zoology
University of Cambridge
Cambridge, England
Karl Sperling
Institut für Humangenetik
Freie Universität Berlin
Berlin, Federal Republic of Germany
ACADEMIC PRESS 1982
A Subsidiary of Harcourt Brace Jovanovich, Publishers
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COPYRIGHT © 1982, BY ACADEMIC PRESS, INC.
ALL RIGHTS RESERVED.
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United Kingdom Edition published by
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Library of Congress Cataloging in Publication Data
Main entry under title:
Premature chromosome condensation.
(Cell biology)
Includes bibliographies and index.
1. Premature chromosome condensation. I. Rao, Potu N.
II. Johnson, Robert T. III. Sperling, Karl. IV. Series.
[DNLM: 1. Cell transformation, Aeoplastic. 2. Chromo-
somes. 3. Gene expression regulation. 4. Neoplasms—
Familial and genetic. QZ 200 P925]
QH600.P73 574.87'322 82-3940
ISBN 0-12-580450-4 AACR2
PRINTED IN THE UNITED STATES OF AMERICA
82 83 84 85 9 8 7 6 5 4 3 2 1
List of Contributors
Numbers in parentheses indicate the pages on which the authors1 contributions begin.
RAMESH C. ADLAKHA (233), Department of Developmental Therapeutics,
University of Texas System Cancer Center, M. D. Anderson Hospital
and Tumor Institute, Houston, Texas 77030
BERND BEEK1 (113), MRC Clinical and Population Cytogenetic Unit, Western
General Hospital, Edinburgh EH4 2XU, Scotland
A. R. S. COLLINS (253), Cancer Research Campaign, Mammalian Cell DNA
Repair Group, Department of Zoology, University of Cambridge, Cam-
bridge CB2 3EJ, England
ALAIN DELEENER (159), Faculteit der Wetenschappen Algemene, Universiteit
Brüssel, B-1050 Brussels, Belgium
HELEN L. DRWINGA (99), Institute for Cancer Research, Fox Chase Cancer
Center, Philadelphia, Pennsylvania 19111
DENES DUDITS (359), Institute of Genetics, Biological Research Center, Hunga-
rian Academy of Sciences, Szeged, Hungary
GYULA HADLACZKY (359), Institute of Genetics, Biological Research Center,
Hungarian Academy of Sciences, Szeged, Hungary
WAHEEB K. HENEEN 2 (79, 131), Institute of Genetics, University of Lund, S-223
62, Lund, Sweden
'Present address: Institut für Genetik, Freie Universität Berlin, D-1000 Berlin 33, Federal Republic
of Germany.
2Present address: Department of Crop Genetics and Breeding, The Swedish University of Agricultural
Sciences, S-268 00 Svalöv, Sweden
xi
xii List of Contributors
JOSEPH HIRSCHBERG (173), Department of Genetics, The Hebrew University of
Jerusalem, Jerusalem 91904, Israel
WALTER N. HITTELMAN (309), Department of Developmental Therapeutics,
University of Texas System Cancer Center, M. D. Anderson Hospital
and Tumor Institute, and University of Texas Graduate School of
Biomedical Sciences, Houston, Texas 77030
ROBERT T. JOHNSON (253), Department of Zoology, University of Cambridge,
Cambridge, England
MENASHE MARCUS (173), Department of Genetics, The Hebrew University of
Jerusalem, Jerusalem 91904, Israel
TOHRU MARUNOUCHI (195), Laboratory of Cell Biology, Mitsubishi-Kasei Insti-
tute of Life Sciences, Machida-shi, Tokyo 194, Japan
SEI-ICHI MATSUI (207), Departments of Genetics and Endocrinology, Roswell
Park Memorial Institute, Buffalo, New York 14263
YOHICHI MATSUMOTO (195), Department of Physiological Chemistry, Faculty of
Pharmaceutical Sciences, University of Tokyo, Hongo, Tokyo 113,
Japan
SHIRO MITA3 (195), Laboratory of Cell Biology, Mitsubishi-Kasei Institute of
Life Sciences, Machida-shi, Tokyo 194, Japan
HEIDEMARIE NEITZEL (159), Institut für Humangenetik, Freie Universität Berlin,
D-1000 Berlin 19, Federal Republic of Germany
GÜNTER OBE (113), Institut für Genetik, Freie Universität Berlin, D-1000
Berlin 33, Federal Republic of Germany
RAJIVA RAMAN (159), Cytogenetics Laboratory, Department of Zoology,
Banaras Hindu University, Varanasi, India
Ροτυ N. RAO (1, 233), Department of Developmental Therapeutics, University
3 Present address: Department of Pharmacology, Keio University School of Medicine, Shinanomachi,
Tokyo, Japan
List of Contributors xiii
of Texas System Cancer Center, M. D. Anderson Hospital and Tumor
Institute, Houston, Texas 77030
DAN RÖHME4 (79, 131), Institute of Genetics, University of Lund, S-223 62,
Lund, Sweden
CHINTAMAN G. SAHASRABUDDHE (233), Department of Molecular Biochemis-
try, University of Texas System Cancer Center, M. D. Anderson Hos-
pital and Tumor Institute, Houston, Texas 77030
AVERY A. SANDBERG (207), Departments of Genetics and Endocrinology, Ros-
well Park Memorial Institute, Buffalo, New York 14263
KARL SPERLING (43, 159), Institut für Humangenetik, Freie Universität Berlin,
D-1000 Berlin 19, Federal Republic of Germany
PRASAD S. SUNKARA (233), Merrill Research Center, Cincinnati, Ohio 45215
LÀSZLO SZABADOS (359), Institute of Genetics, Biological Research Center,
Hungarian Academy of Sciences, Szeged, Hungary
C. A. WALDREN (253), Eleanor Roosevelt Institute for Cancer Research and
Department of Radiology, University of Colorado Health Sciences
Center, Denver, Colorado 80262
HERBERT WEINFELD (207), Departments of Genetics and Endocrinology, Ros-
well Park Memorial Institute, Buffalo, New York 14263
DAVID A. WRIGHT (233), Department of Biology, University of Texas System
Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston,
Texas 77030
HIDEYO YASUDA5 (195), Department of Physiological Chemistry, Faculty of
Pharmaceutical Sciences, University of Tokyo, Hongo, Tokyo 113,
Japan
4Present address: European Molecular Biology Laboratory, Heidelberg 6900, Federal Republic
of Germany
5Present address: Departments of Genetics and Endocrinology, Roswell Park Memorial Institute,
Buffalo, New York 14263
Preface
Since the 1960s the controlled fusion between different cells by means of
Sendai virus and, more recently, polyethylene glycol, has provided biologists
with a powerful tool with which to explore many fundamental problems that
previously were largely inaccessible. One such area is the regulation of the cell
cycle, particularly of DNA synthesis and mitosis. Fusion permitted the ready
construction of cells containing elements from defined and different stages of the
cycle and led directly to the observation and understanding that condensation of
interphase chromatin is associated with asynchrony in the cell. The varied nature
of the condensed interphase structures was explicable in terms of cell cycle
stages, and this knowledge helped to illuminate earlier results from a wide range
of biological materials. The term premature chromosome condensation was used
to describe the phenomenon.
Premature chromosome condensation has since proved to be a fruitful area of
research, bringing together workers from many disciplines. It was therefore
decided that a workshop supported by the International Union against Cancer be
held to review the current status of research and to explore future possibilities,
not least in relation to cancer. The workshop held at the Institut für
Humangenetik, Freie Universität Berlin, on September 6, 1980 provided the
impetus for this volume, which presents a series of detailed reviews of the
contributions that prematurely condensed chromosomes have made to different
areas of research. We hope that these chapters will serve as a stimulus to attract
new investigators to this field.
We hope that the readers, whether working in basic and applied research or in
teaching, will experience the same fascination as the scientists who joined this
meeting: the cytogeneticists, since this experimental approach makes it possible
to look at the chromosomes as discrete structures at any stage in a cell's life and
thereby obtain deeper insight into the chromosomal architecture and its regular
conformational and functional changes during the cell cycle; the cell biologists,
since this system provides a deeper understanding of the cascade of events
associated with the normal process of mitosis and meiosis in animals and in
plants; the biochemist, who makes use of this system to approach the fundamen-
xv
xvi Preface
tal physiological and molecular mechanisms that control those processes. Inves-
tigators in the field of mutation research will profit from the ability to analyze
chromosomes shortly after induction of DNA damage and to follow the course of
chromosome repair not only in G,, G , or G cells but also in differentiated cells
2 ()
which do not normally divide. This approach was also sought to determine the
efficiency of chemotherapy in patients with malignancy. Quite unexpectedly it
led to the development of a most valuable method for predicting the clinical
course of patients with leukemia, which can now serve as a basis for individual
therapeutic strategy. Thus, premature chromosome condensation becomes
another tool for the clinician in the treatment of human malignancies.
We gratefully acknowledge the financial sponsorship for this workshop pro-
vided primarily by the "International Cancer Research Workshop Programme"
(ICREW), Geneva, and the generous support by the Freie Universität Berlin. We
are indebted to the members of the Institut für Humangenetik for their help in
making the local arrangements for the workshop. Our special thanks go to the
contributing authors for their enthusiasm and willingness to write about their
work. We are extremely grateful to Walter J. Pagel for his assistance in editing
this volume.
Potu N. Rao
Robert T. Johnson
Karl Sperling
1
The Phenomenon of Premature
Chromosome Condensation
Potu N. Rao
I. Introduction 2
II. The Phenomeon of Premature Chromosome Condensation . . .. 2
A. Mitotic Factors Have No Species Specificity 6
B. Visualization of Chromosomes from Differentiated Cells . .. 6
C. Some Generalizations 8
III. Historical Background 10
IV. Structure of Prematurely Condensed Chromosomes (PCC) . . .. 12
A. G, PCC 12
B. S PCC 15
C. G, PCC 18
V. The Fate and Consequences of Premature Chromosome Condensation 21
A. Template Activity of Prematurely Condensed Chromosomes 21
B. Initiation of DNA Synthesis in G, PCC 21
C. Integration or Exclusion of Prematurely Condensed Chromosomes
in Hybrid Cells 23
D. Effect of an Interphase Component on Metaphase Chromosomes 29
VI. Factors Involved in the Induction of Premature Chromosome
Condensation 30
A. Effect of Anions and Cations on the Induction of Premature
Chromosome Condensation 31
Β. Involvement of Proteins in the Induction of Premature
Chromosome Condensation 32
VII. Differences between Prematurely Condensed Chromosomes and
Metaphase Chromosomes 35
VIII. Summary 37
References 37
1
PREMATURE CHROMOSOME CONDENSATION
Copyright © 1982 by Academic Press, Inc.
All rights of reproduction in any form reserved.
ISBN 0-12-580450-4
2 Potu Ν. Rao
I. INTRODUCTION
The discovery that mammalian cells in culture can be readily fused by using
uv-inactivated Sendai virus (Okada et al., 1957; Harris, 1965) has opened up
new frontiers in cell biology. Some of the most notable advances made by the use
of the cell fusion technique include the mapping of the human genome, the
development of the hybridoma technique, and the discovery of the phenomenon
of premature chromosome condensation. The objective of this chapter is to
review the phenomenon of premature chromosome condensation, its discovery,
its historical background, and the structural aspects of its products, the prema-
turely condensed chromosomes (PCC).
In plant and animal cells, chromosomes can be visualized as discrete cytologi-
cal entities only for a brief time during the life cycle, i.e., either during mitosis or
meoisis. Throughout the entire intermitotic period, which varies from 10 to 40 hr
for animal cells in culture, the chromosomes remain diffuse and indistinguishable
in the nucleus. The DNA molecule, complexed with histone and non-histone
proteins, undergoes several orders of coiling, giving rise to chromatin fibers of
about 230 Â in diameter; these are commonly observed by electron microscopy
in interphase nuclei or isolated metaphase chromosomes (Ris, 1956; Gay, 1956;
Abuelo and Moore, 1969; DuPraw, 1968). Such fibers are tightly packed into
chromosomes at the time of cell division, a process that facilitates the equal
distribution of genetic material between the two daughter cells.
It has been almost a century since the chromosome cycle in plant and animal
cells was first described (Flemming, 1880), yet key questions still remain unan-
swered: (1) How is the presumably ordered yet dispersed interphase chromatin
organized into the dense chromosome bodies? (2) What is the nature of the
intracellular signals that trigger dissolution of the nuclear membrane and conden-
sation of chromosomes? Since chromosome condensation is invariably as-
sociated with mitosis, an understanding of this process might give us some
insight into the regulation of cell division in normal and malignant cells.
In this chapter I also discuss the attempts to promote condensation of chromo-
somes experimentally, some of the factors that may be involved in this process,
the fate and consequences of premature chromosome condensation, and the role
of polyamines, cations, and proteins in the induction of premature chromosome
condensation.
II. THE PHENOMENON OF PREMATURE CHROMOSOME
CONDENSATION
Initially we set out to study the pattern of mitotic accumulation among binuc-
leate HeLa cells, which were formed by the fusion of S phase cells (prelabeled
