Table Of ContentChromatin Structure
and Function
Levels of Organization
and Cell Function
PartB
NATO ADVANCED STUDY INSTITUTES SERIES
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Volume 210 - Chromatin Structure and Function:
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Volume 21 b - Chromatin Structure and Function:
Levels of Organization and Cell Function
edited by Claudio A. Nicolini
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Chromatin Structure
and Function
Levels of Organization
and Cell Function
PartB
Edited by
Claudio A. Nicolini
Temple University
Philadelphia, PennsylvaniQ
PLENUM PRESS. NEW YORK AND LONDON
Published in cooperation with NATO Scientific Affairs Division
Library of Congress Cataloging in Publication Data
Nato Advanced Study Institute, Erice, Italy, 1978.
Chromatin structure and function.
(NATO advanced study institutes series: Series A, Life sciences; v. 21)
Includes bibliographical references and indexes.
CONTENTS: pt. A. Molecular and cellular biophysical methods. - pt. B. Levels
of organization and cell function.
1. Chromatin - Congresses. 2. Carcinogenesis - Congresses. 1. Nicolini, Claudio A.
II. Title. III. Series.
QH599.N37 1978 574.8'732 78-24268
ISBN-13: 978-1-4684-0978-9 e-ISBN-13: 978-1-4684-0976-5
DOl: 10.1007/978-1-4684-0976-5
Second half of the Proceedings of the NATO Advanced Study
Institute held at Erice, Italy, April 12-26, 1978
© 1979 Plenum Press, New York
Softcover reprint of the hardcover 1st edition 1979
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N.Y. 10011
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical, photocopying, microfJ1ming,
recording, or otherwise, without written permission from the Publisher
To
My Parents
and
Uncle Luigi
PREFACE
This volume is the second part of the book on "Chromatin
Structure and Function", which resulted from a NATO-Advanced
Study Institute held at Erice during April 1978. In addition to
giving an updated and detailed description of various levels of
chromatin organization, i.e. octamers, nucleosomes, multimers,
solenoid and higher order fibers (including the most recent, yet
unpublished, findings), it focuses, in a tutorial and organic
format, on the possible mechanisms controlling transcription and
on the basic biological phenomena (either genetic or epigenetic)
related to cell aging, cell cycle, differentiation, transformation
and chemical carcinogenesis. The most significant (sometime
spirite~ discussion sessions have been included at the end of
every section. Their clarifying nature is further supported by
the final section (V), which summarizes and reviews the current
state of the art on the genetic apparatus and its constituents.
All chapters have selected up-to-date references; quite a few
have an extensive bibliography both in terms of basic reference
books and most recent findings.
A few chapters, dealing at the level of intact cell and/or in
classical genetic terms, with aging, differentiation and neoplastic
transformation, have been included to furnish a more comprehensive
view of fundamental cell functions directly and indirectly related
to the structure and function of the genetic apparatus.
At the same time it is hoped that interested students and
investigators will find in the chapters of this volume the
necessary and stimulating introduction to the wide variety of
fundamental mechanism and phenomena occurring in higher eukaryotes.
Claudio Nicolini
vii
CONTENTS OF PART A
Introduction xi
C. Nicolini
SECTION I: WHAT IS THE CHROMATIN?
Properties and Composition of
Isolat.ed Chromatin 3
J. Bonner
Expressed and Nonexpressed Portions of
the Genome: Their Separation and
Their Characterization • • • • • 15
J. Bonner
Discussion 25
SECTION II: PHYSICAL, CHEMICAL AND
BIOLOGICAL TECHNIQUES FOR STUDYInG NUCLEOSOME,
CHROMATIN, CHROMOSOME AND NUCLEI
Electron Microscopy: A Tool for
Visualizing Chromatin 31
A. L. Olins
Transcriptional Control of
Native Chromatin 41
R. S. Gilmour
Circular Dichroism of DNA, Protein
and Chromatin • • • • • • 67
G. D. Fasman
Important Hydrodynamic and Spectroscopic
Techniques in the Field of Chromatin
Structure • • • • • • • • • • • • 109
D. E. Olins
ix
x CONTENTSOFPARTA
Preparation and Analysis of Core Particles
and Nucleosomes: A Conveinient Method
For Studying the Protein Composition
of Nucleosomes Using Protamine-Release
into Triton-Acid-Urea Gels ••• • • • 125
B. R. Shaw and R. G. Richards
The Interaction of Histones with
DNA: Equilibrium Binding Studies 137
D. R. Burton, M. J. Butler, J. E. Hyde,
D. Phillips, C. J. Skidmore and I. O. Walker
Nucleosome Shape and Structure in
Solution from Flow Birefringence 167
R. E. Harrington
Scattering and Diffraction by
Neutrons and X-rays in the
Study of Chromatin 187
J. F. Pardon
Nuclear Magnetic Resonance Studies
of Nucleic Acids and Proteins 217
P. O. P. Ts'o and L.-S. Kan
Techniques for Cytochemical Studies
of the Nucleus and its
Substructures 251
T. Caspers son
Chromatin Study in Situ: I. Image Analysis 265
F. Kendall, F. Beltrame and C. Nicolini
Chromatin Study in Situ: II. Static
and Flow Microfluorimetry 293
C. Nicolini, S. Parodi, S. Lessin,
A. Belmont, S. Abraham, S. Zietz and M. Grattarola
Chromatin Study in Situ: III. Differential
Effects of Feulgen Hydrolysis • 323
W. A. Linden, S. M. Fang, S. Zietz and
C. Nicolini
Scanning and Flow Photometry of Chromosomes • • • • • • •• 341
M. L. Mendelsohn
Discussion 357
Index • • • • xxi
CONTENTS OF PART B
Introduction xiii
C. Nicolini
SECTION III: VARIOUS LEVELS OF
CHROMATIN ORGANIZATION AND MECHANISMS
FOR TRANSCRIPTIONAL CONTROL
Histones Assembly and Their Structural
Role for Nucleosome Core 371
N. M. Maraldi, S. Capitani, L. Cocco and
F. A. Manzoli
Nuclease Digestion and the Structure
of Chromatin •••••• 389
K. E. Van Holde, J. R. Allen, J. Corden, D. Lohr,
K. Tatchell and W. O. Weischet
Reconstitution of Nucleosomes • • • 413
K. Tatchell and K. E. Van Holde
Conformation of Polynucleosomes in Low
Ionic Strength Solution 427
B. R. Shaw and K. S. Schmitz
Chromatin Structure: Relation of Nucleosomes
of DNA Sequences • • • • • • • • • 441
A. Prunell
Histone Complexes, Nucleosomes, Chromatin and
Cell-Cycle Dependent Modification
of Histones . • . . . . . . . . . . 451
H. W. E. Rattle, G. G. Kneale, J. P. Baldwin,
H. R. Matthews, C. Crane-Robinson, P. D. Cary,
B. G. Carpenter, P. Suau and E. M. Bradbury
xi
xii CONTENTS OF PART B
Evidence for Superstructures of Wet
Chromatin • • • • • • 515
S. Basu
Chromatin Fractionation and the Properties
of Transcriptionally Active Regions
of Chromatin • • • • • • • • • • • 541
J. Gottesfeld
Chromatin Reconstitution and Non-Histone
Proteins 561
R. S. Gilmour
Discussion 593
SECTION IV: STRUCTURE-FUNCTION OF THE
GENETIC APPARATUS AND CELL CYCLE,
AGING, NEOPLASTIC TRANSFORMATION,
DIFFERENTIATION, CHEMICAL CARCINOGENESIS
The Structure and Function of Chromatin
in Lower Eukaryotes 599
K. E. Davies and I. O. Walker
Chromatin Structure from Angstrom to Micorn
Levels, and Its Relationship to
Mammalian Cell Proliferation 613
C. Nicolini
Chromatin Pattern in Situ: Dependence upon
Cell Cycle, Preimplantation and
Development, and Cellular Aging
in Vitro ••••••••••• 667
W. Sawicki
Neoplastic Transformation: The Relevance of
in Vitro Studies for the Understanding
of Tumor Pathenogenesis and Neoplastic
Growth • • • • • • . • • • . • • • • . 683
L. A. Smets
Cell Differentiation and Malignancy
in Leukemia • • • • • • • 705
L. Sachs
Cellular Morphometry in Transformation,
Differentiation and Aging • • 721
S. Parodi, G. Brambilla, F. Beltrame, S. Lessin
and C. A. Nicolini
