Characterization of the Human Recombination Activating Gene 1 (RAG1) and RAG2 Promoter Regions

Characterization of the Human Recombination Activating Gene 1 (RAG1) and RAG2 Promoter Regions
Author: Ali Akbar Zarrin
Publisher:
Total Pages: 0
Release: 2000
Genre:
ISBN:

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In all vertebrates, the genes encoding Antigen Receptor (AgR) variable (V) regions are assembled during B and T cell development from gene segments termed V, (D), and J through the process of V(D)J recombination. V(D)J recombination is central for the successful function and development of B and T cell lymphocytes. Recombination Activating Gene 1 (RAG1) and RAG2 are the essential and tissue specific components of V(D)J recombination. Their precise pattern of expression during B and T cell development determines where and when V(D)J recombination occurs. This thesis describes studies on the regulation of RAG1 and RAG2 at the transcription level. In chapter 2, I have characterized the exon 1 and promoter region of human RAG1. I show that a single promoter region is used for RAG1 transcription. In chapter 3, I describe the isolation and characterization of the human RAG2 promoter regions. The major transcription initiation site (exon 1A) of human RAG2 is located 3.9 kb upstream of the coding exon while the less frequently used transcription initiation site (exon 1B) is only 0.7 kb upstream of the coding exon. Two alternate promoters upstream of exon1A or 1B drive the transcription of RAG2 gene. Using transient assays, I show that promoter regions of human RAG1 and RAG2 are active in both lymphoid and non-lymphoid cell lines. Chapter 4 describes an approach that I took to study the tissue-specific regulation of RAG genes. I stably transfected a 65 kb P1 phage genomic clone containing RAG1, RAG2 and their surrounding sequences, into the human cervical carcinoma cell line, HeLa (RAG1-, RAG2-) and the mouse pre-B cell line, NFS-70 (RAG1+, RAG2+). I found consistent expression of the human RAG1 gene in multiple clones of NFS-70 cell line while no RAG1 expression was detected in HeLa clones suggesting that this genomic clone contains some of the sequences required for the tissue-specific regulation of RAG1. In chapter 5, I describe the second approach to isolate the potential regulatory elements by subcloning fragments of the P1 phage into the RAG1 promoter construct and subsequently looking for enhancer or silencer activity. I located a 1425 bp within the 3' untranslated region of the RAG1 gene with enhancer activity. In chapter 6, I describe the subcloning and sequencing a 7.3 kilobase of the 5' flanking region of human RAG2 to look for potential conserved regulatory elements between human and mouse species. In Chapter 7, models of RAG1/RAG2 promoters, and their tissue-specific transcriptional regulation are discussed.

Molecular Biology of B Cells

Molecular Biology of B Cells
Author: Tasuku Honjo
Publisher: Academic Press
Total Pages: 0
Release: 2014-12-22
Genre: Science
ISBN: 9780123979339

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Molecular Biology of B Cells, Second Edition is a comprehensive reference to how B cells are generated, selected, activated and engaged in antibody production. All of these developmental and stimulatory processes are described in molecular, immunological, and genetic terms to give a clear understanding of complex phenotypes. Molecular Biology of B Cells, Second Edition offers an integrated view of all aspects of B cells to produce a normal immune response as a constant, and the molecular basis of numerous diseases due to B cell abnormality. The new edition continues its success with updated research on microRNAs in B cell development and immunity, new developments in understanding lymphoma biology, and therapeutic targeting of B cells for clinical application. With updated research and continued comprehensive coverage of all aspects of B cell biology, Molecular Biology of B Cells, Second Edition is the definitive resource, vital for researchers across molecular biology, immunology and genetics.

Immunoglobulin Genes

Immunoglobulin Genes
Author: Tasuku Honjo
Publisher: Elsevier
Total Pages: 464
Release: 1995-09-27
Genre: Medical
ISBN: 0080534511

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The immunoglobulin gene complex is responsible for generating an extraordinarily wide range of antibodies, each possessing a unique antigen specificity. The Second Edition of Immunoglobulin Genes brings the reader up to date with the rapid progress in our understanding of this system. Firmly established as the definitive book on the topic, it provides a fully comprehensive account of the organization, function, rearrangement and expression of these and related genes. Since the publication of the first edition, the work has been extensively revised and updated to cover new research data on human immunoglobulin genes as well as mammalian and lower vertebrate systems. New chapters include "Somatic mutation and B cell Maturation", "Antigen receptors" and "B lymphocyte tolerance in the mouse." For the immunologist, there are detailed descriptions of the molecular mechanisms which generate this diverse range of antibodies and the tight regulation of the genes encoding them. Molecular biologists will find discussions of important phenomena including gene rearrangement, differential splicing and deletion in the context of immunoglobulin genes. The excitement of research on immunoglobulin genes is conveyed by the range and complexity of issues discussed here; the book contains invaluable information for clinicians and research scientists with an interest in the immune response. * Conveys the excitement of research on immunoglobulin genes by the range and complexity of issues * Contains invaluable information for clinicians and research scientists with an interest in the immune response * For the Immunologist * Detailed descriptions of the molecular mechanisms generating this diverse range of antibodies and the regulation of genes encoding them * For the Molecular Biologist * Discusses important phenomena including gene rearrangement, differential splicing, and deletion in the context of immunoglobulin genes.

Molecular Analysis of B Lymphocyte Development and Activation

Molecular Analysis of B Lymphocyte Development and Activation
Author: Harinder Singh
Publisher: Springer Science & Business Media
Total Pages: 261
Release: 2006-03-30
Genre: Medical
ISBN: 3540263632

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The B lymphocyte lineage represents an important paradigm for exploring the molecular mechanisms underlying cell fate specification, differentiation and cellular activation. In the past five years, major advances have been achieved in our understanding of the transcriptional control of early B cell development and terminal plasma cell differentiation. In addition new insights became available for the processes of B cell activation, class switch recombination and somatic hypermutation. Many of the new findings and their implications for a molecular understanding of B cell biology in particular and cell differentiation in general are covered in this volume.

Mobile DNA III

Mobile DNA III
Author: Michael Chandler
Publisher: John Wiley & Sons
Total Pages: 1321
Release: 2020-07-24
Genre: Science
ISBN: 1555819214

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An exploration of the raw power of genetic material to refashion itself to any purpose... Virtually all organisms contain multiple mobile DNAs that can move from place to place, and in some organisms, mobile DNA elements make up a significant portion of the genome. Mobile DNA III provides a comprehensive review of recent research, including findings suggesting the important role that mobile elements play in genome evolution and stability. Editor-in-Chief Nancy L. Craig assembled a team of multidisciplinary experts to develop this cutting-edge resource that covers the specific molecular mechanisms involved in recombination, including a detailed structural analysis of the enzymes responsible presents a detailed account of the many different recombination systems that can rearrange genomes examines the tremendous impact of mobile DNA in virtually all organisms Mobile DNA III is valuable as an in-depth supplemental reading for upper level life sciences students and as a reference for investigators exploring new biological systems. Biomedical researchers will find documentation of recent advances in understanding immune-antigen conflict between host and pathogen. It introduces biotechnicians to amazing tools for in vivo control of designer DNAs. It allows specialists to pick and choose advanced reviews of specific elements and to be drawn in by unexpected parallels and contrasts among the elements in diverse organisms. Mobile DNA III provides the most lucid reviews of these complex topics available anywhere.