Chromosomal Proteins and their Role in the Regulation of Gene Expression PDF
Preview Chromosomal Proteins and their Role in the Regulation of Gene Expression
Colloquium Organizing Committee Gary Stein, Chairman Lewis Kleinsmith, Co-chairman Peter Cerutti Rusty Mans Philip Laipis Eugene Sander Proceedings of the Florida Colloquium on Molecular Biology March 13-14, 1975 Sponsored by the Department of Biochemistry University of Florida Gainesville, Florida Chromosomal Proteins and Their Role in the Regulation of Gene Expression Edited by GARY S. STEIN Department of Biochemistry The University of Florida Gainesville, Florida LEWIS J. KLEINSmiTH Department of Zoology The University of Michigan Ann Arbor, Michigan ® Academic Press, Inc. New York San Francisco London 1975 A Subsidiary of Harcourt Brace Jovanovich Publishers COPYRIGHT © 1975, BY ACADEMIC PRESS, INC. ALL RIGHTS RESERVED. NO PART OF THIS PUBLICATION MAY BE REPRODUCED OR TRANSMITTED IN ANY FORM OR BY ANY MEANS, ELECTRONIC OR MECHANICAL, INCLUDING PHOTOCOPY, RECORDING, OR ANY INFORMATION STORAGE AND RETRIEVAL SYSTEM, WITHOUT PERMISSION IN WRITING FROM THE PUBLISHER. ACADEMIC PRESS, INC. Ill Fifth Avenue, New York, New York 10003 United Kingdom Edition published by ACADEMIC PRESS, INC. (LONDON) LTD. 24/28 Oval Road, London NW1 Library of Congress Cataloging in Publication Data Florida Colloquim on Molecular Biology, University of Florida, Gainesville, 1975. Chromosomal proteins and their role in the regulation of gene expression. "Proceedings of the Florida Colloquim on Molecular Biology, March 13-14, 1975." Includes bibliographies and index. 1. Molecular biology-Congresses. I. Stein, Gary S. II. Kleinsmith, Lewis J. III. Florida. University, Gainesville. Dept. of Biochemistry. [DNLM: 1. Genetics, Biochemical-Congresses. 2. Chromosomes- Physiology-Congresses. 3. Chromatin-Physiology- Congresses. 4. Proteins-Physiology-Congresses. QH450F636c 1975] QH506.F55 1975 574.8'8 75-22181 ISBN0-12-664750-X PRINTED IN THE UNITED STATES OF AMERICA Contributors Vincent G. Allfrey, The Rockefeller University, New York, New York 10021 Raghuveera Ballal, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 C. Stuart Baxter, Department of Pathology, University of Florida, College of Medicine, Gainesville, Florida 32610 James Bonner, Division of Biology, California Institute of Technology, Pasadena, California 91125 Harris Busch, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Rose K. Busch, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Paul Byvoet, Department of Pathology, University of Florida, College of Medicine, Gainesville, Florida 32610 Roger Chalkley, Department of Biochemistry, The University of Iowa, Iowa City, Iowa 52242 Edward Ezrailson, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Carl M. Feldherr (Session Chairman), Department of Pathology, University of Florida, Gainesville, Florida 32610 Gary Felsenfeld, Laboratory of Molecular Biology, National Institute of Arthritis, Metabolism, and Digestive Diseases, Bethesda, Maryland 20014 R. Stewart Gilmour, The Beatson Institute for Cancer Research, Glasgow, Scotland G3 6UD Ira L. Goldknopf, Nuclear Protein and Tumor By-Products Laboratories, VII CONTRIBUTORS Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Joel Gottesfeld, Division of Biology, California Institute of Technology, Pasadena, California 91125 Daryl K. Granner, Department of Biochemistry, The University of Iowa, Iowa City, Iowa 52242 Sidney R. Grimes, Department of Biochemistry, University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston, Texas 77025 Lubomir S. Hnilica, Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee 37232 Philip Hohmann, Biomedical Research Group, H-9, Los Alamos Scientific Laboratory, Los Alamos, New Mexico 87544 Gwen Hord, Department of Biochemistry, University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston, Texas 77025 Akira Inoue, The Rockefeller University, New York, New York 10021 Vaughn Jackson, Department of Biochemistry, The University of Iowa, Iowa City, Iowa 52242 Edward M. Johnson, The Rockefeller University, New York, New York 10021 Diane Kent, Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125 Lewis J. Kleinsmith (Session Chairman), Department of Zoology, University of Michigan, Ann Arbor, Michigan 48104 Nina C. Kostraba, Division of Cell and Molecular Biology, State University of New York at Buffalo, Buffalo, New York 14214 Thomas A. Langan, Department of Pharmacology, University of Colorado Medical School, Denver, Colorado 80220 Rusty J. Mans (Session Chairman), Department of Biochemistry, University of Florida, Gainesville, Florida 32610 Marvin L. Meistrich, Department of Experimental Radiotherapy, University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston, Texas 77025 Mark O. J. Olson, Nuclear Protein and Tumor By-Products Laboratories, viii CONTRIBUTORS Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 William Park, Department of Biochemistry, University of Florida, Gainesville, Florida 32610 Gordhan L Patel, Department of Zoology, University of Georgia, Athens, Georgia 30602 John Paul, The Beatson Institute for Cancer Research, Glasgow, Scotland G3 6UD George M. Pikier, Department of Molecular Medicine, Mayo Clinic, Rochester, Minnesota 55901 Robert D. Platz, Department of Biochemistry, University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Houston, Texas 77025 Archie W. Prestayko, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Michael Ross, Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125 Barbara Sollner-Webb, Laboratory of Molecular Biology, National Institute of Arthritis, Metabolism, and Digestive Diseases, Bethesda, Maryland 20014 Thomas C. Spelsberg, Department of Molecular Medicine, Mayo Clinic, Rochester, Minnesota 55901 Gary S. Stein (Session Chairman), Department of Biochemistry, University of Florida, Gainesville, Florida 32610 Janet L. Stein, Department of Biochemistry, University of Florida, Gainesville, Florida 32610 Charles W. Taylor, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Terry L. Thomas, Department of Zoology, University of Georgia, Athens, Georgia 30602 Cary Thrall, Department of Biochemistry, University of Florida, Gainesville, Florida 32610 Tung Yue Wang, Division of Cell and Molecular Biology, State University of New York at Buffalo, Buffalo, New York 14214 IX CONTRIBUTORS Robert Webster, Department of Molecular Medicine, Mayo Clinic, Rochester, Minnesota 55901 Lynn C. Yeoman, Nuclear Protein and Tumor By-Products Laboratories, Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77025 Preface It was not long ago that considerations of the eukaryotic genome focused almost entirely on DNA, whereas chromosomal proteins were only briefly men tioned or at times even ignored. In recent years the situation has changed dra matically. Intensive efforts have now been directed toward examining the proteins associated with the genome, with initial attention being focused on the histones followed by a growing concern with the nonhistone chromosomal proteins. It is becoming increasingly apparent that these classes of protein may dictate im portant structural as well as transcriptional properties of the genetic material. The existence of histones in nuclei of eukaryotic cells was documented prior to the turn of the century, and an involvement of these basic chromosomal proteins in the regulation of gene expression was postulated as early as three decades ago. However, it has only been during the past 15 years that intensive experimental efforts employing metabolic and structural probes have yielded con vincing evidence that histones play important roles in both restricting the tran scriptional capacity of DNA and in the packaging of the genetic information. Despite the level to which our understanding of the structural and functional properties of the histones has progressed, many important problems remain to be resolved. The precise manner in which histones interact with other genome com ponents requires clarification. The interrelationship of histone synthesis and DNA replication must be further elucidated. The distribution of histones along active and inactive regions of the DNA molecule is not known. Answers to these questions and many others are undoubtedly forthcoming. Although a structural and regulatory role for the histones has been clearly documented, it has become increasingly apparent that these proteins lack the specificity for recognition of individual genetic loci. As a result, attention has recently turned to the nonhistone protein fraction as a potential source of specific regulators of gene readout, i.e., molecules capable of interacting with defined DNA sequence in such a manner as to render them more or less transcribable. These proteins originally were described as early as the 1940s, but their insolu bility and tendency to form aggregates has presented a major obstacle to their XI PREFACE characterization. This problem was partially circumvented in the late 1960s with the development of SDS polyacrylamide gel electrophoretic fractionation methods, which afforded an opportunity for separating the nonhistone chromo somal proteins in a soluble (although denatured) state. As a result, it has been possible to demonstrate in numerous biological systems that variations in the composition and metabolism of nonhistone chromosomal proteins are correlated with modifications in gene expression. Some of the nonhistone proteins have also been shown to be capable of specific binding to DNA. Both of these types of observations support the notion that these proteins are in some way involved in the regulation of transcription. More direct evidence has come from chromatin reconstitution experiments, which have demonstrated that components of the nonhistone chromosomal proteins mediate control of transcription of tissue- specific and cell cycle stage-specific genes. Although it is undeniable that we have made definitive inroads toward elucidating the involvement of nonhistone chromosomal proteins in the regulation of transcription, we have merely un covered the first set of clues to an extremely complex problem. Perhaps the most obvious problem is identifying which among the highly heterogeneous nonhistone chromosomal proteins are the specific regulatory proteins. Another series of prob lems concerns the distribution of the nonhistone chromosomal proteins in chromatin and the mechanism by which they render genetic sequences transcribable. This book resulted from the 1975 Florida Colloquium on Molecular Biology, the topic of which was "Chromosomal Proteins and Their Role in the Regulation of Gene Expression." The colloquium, and hence the book, considered many current approaches for studying the structural and functional properties of chromosomal proteins, particularly as they relate to the control of transcription. Predicated upon the rapid progress in this area during the past several years and the recent development of high resolution probes for further work, it is reasonable to anticipate that problems previously unresolvable can now be mean ingfully pursued. The editors are indebted to Janet Stein for her editorial assistance and to Bonnie Cooper for typing the manuscripts. XII REGULATION OF HISTONE GENE TRANSCRIPTION DURING THE CELL CYCLE BY NONHISTONE CHROMOSOMAL PROTEINS Gary Stein, Janet Stein, Cary Thrall and William Park Department of Biochemistry University of Florida Gainesville, Florida Abstract We have examined the regulation of histone gene expres sion during the cell cycle and the role of nonhistone chromo somal proteins in such regulation. Adenylic acid residues were added to the 3f-OH ends of histone messenger RNAs, and using RNA-dependent DNA polymerase, a complementary DNA (cDNA) was synthesized. This high resolution probe has been utilized to demonstrate that: a. Histone messenger RNA sequences are present in the polyribosomal RNA of S phase but not of G^ cells; b. Transcription ±n vitro of histone messenger RNA sequences is restricted to the S phase of the cell cycle; and c. Nonhis tone chromosomal proteins are responsible for regulating the transcription of those regions of the genome which contain the information for the synthesis of histones. The cell cycle of continuously-dividing cells provides an effective model system for studying the regulation of gene expression. Many of the complex and interdependent biochemical events occurring throughout the cell cycle require the differ ential elaboration of information contained within the genome (1,2). The onset of DNA replication and mitosis are both essen tial events for which RNA synthesis is an absolute requirement, indicating that their regulation resides, at least in part, at the transcriptional level. An important property of the cell cycle is that it offers the opportunity to study the mechanism by which the transient expression of genes is regulated. This is in contrast to the control of genes which are expressed on a more permanent basis, as in differentiated systems such as erythroid cells which are committed to the synthesis of hemo globin. Several lines of evidence suggest that nonhistone chromo somal proteins play a key role in the regulation of gene expres sion in general (1,3-7) and, specifically, in the control of 1
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