Poly(ADP-ribose) Polymerase Methods and Protocols
Poly (ADP-ribose) Polymerases (PARPs) are abundant and ubiquitous proteins that regulate crucial processes of the cell cycle, DNA repair, genomic stability, and transcriptional regulation. Being involved in basic cell functions, PARPs mediate rapid
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IN
MOLECULAR BIOLOGY™
Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: i http://www.springer.com/series/7651
Poly(ADP-ribose) Polymerase Methods and Protocols Edited by
Alexei V. Tulin Epigenetics and Progenitor Cells Program, Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, PA, USA
Editor Alexei V. Tulin Epigenetics and Progenitor Cells Program Cancer Biology Program Fox Chase Cancer Center Philadelphia, Pennsylvania USA [email protected]
ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-61779-269-4 e-ISBN 978-1-61779-270-0 DOI 10.1007/978-1-61779-270-0 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: 2011933600 © Springer Science+Business Media, LLC 2011 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. Cover illustration: Detection of poly(ADP-ribose) at the spindle pole of mitotic poly(ADP-ribose) Glycohydrolase knock down Hela cell by immunofluorescence microscopy (Jean-Christophe Amé). PAR (in red) was detected using the 10H anti-PAR antibody (IgG3kappa, T. Sugimura). Alpha tubulin (in green) was detected using an anti-alpha-tubulin antibody (DM1a, IgG1kappa, Sigma). DNA was detected by incubating the cell with DAPI. The image was obtained following 3D deconvolution of a 70 images stack in the three colors. All the images were reassembled to obtain a single image (Volocity, Improvision). Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
Preface Our understanding of how Poly(ADP-ribose) polymerases (PARPs) function has reached a new level. It has become clear that these abundant and ubiquitous proteins regulate crucial processes of the cell cycle, DNA repair, genomic stability, and transcriptional regulation. Being involved in basic cell functions, PARPs mediate rapid responses to environmental stress, infection, nutrition changes, and hormonal signals. PARP research has gained tremendous importance for advancement of cancer treatments following a recent discovery that tumor growth can be suppressed by PARP inhibitors in people with a BRCA1 or BRCA2 mutations, which are typical in breast, ovarian, and prostate cancer. The basic enzymatic reactions catalyzed by PARPs involve transferring ADP-ribose moiety from NAD either to a protein
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