Single molecule analysis of c-myb alternative splicing reveals novel classifiers for precursor B-ALL
The c-Myb transcription factor, a key regulator of proliferation and differentiation in hematopoietic and other cell types, has an N-terminal DNA binding domain and a large C-terminal domain responsible for transcriptional activation, negative regulation and determining target gene specificity. Over...
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Published in | PloS one Vol. 6; no. 8; p. e22880 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
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11.08.2011
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Abstract | The c-Myb transcription factor, a key regulator of proliferation and differentiation in hematopoietic and other cell types, has an N-terminal DNA binding domain and a large C-terminal domain responsible for transcriptional activation, negative regulation and determining target gene specificity. Overexpression and rearrangement of the c-myb gene (MYB) has been reported in some patients with leukemias and other types of cancers, implicating activated alleles of c-myb in the development of human tumors. Alternative RNA splicing can produce variants of c-myb with qualitatively distinct transcriptional activities that may be involved in transformation and leukemogenesis. Here, by performing a detailed, single molecule assay we found that c-myb alternative RNA splicing was elevated and much more complex in leukemia samples than in cell lines or CD34+ hematopoietic progenitor cells from normal donors. The results revealed that leukemia samples express more than 60 different c-myb splice variants, most of which have multiple alternative splicing events and were not detectable by conventional microarray or PCR approaches. For example, the single molecule assay detected 21 and 22 splice variants containing the 9B and 9S exons, respectively, most of which encoded unexpected variant forms of c-Myb protein. Furthermore, the detailed analysis identified some splice variants whose expression correlated with poor survival in a small cohort of precursor B-ALL samples. Our findings indicate that single molecule assays can reveal complexities in c-myb alternative splicing that have potential as novel biomarkers and could help explain the role of c-Myb variants in the development of human leukemia. |
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AbstractList | The c-Myb transcription factor, a key regulator of proliferation and differentiation in hematopoietic and other cell types, has an N-terminal DNA binding domain and a large C-terminal domain responsible for transcriptional activation, negative regulation and determining target gene specificity. Overexpression and rearrangement of the c-myb gene (MYB) has been reported in some patients with leukemias and other types of cancers, implicating activated alleles of c-myb in the development of human tumors. Alternative RNA splicing can produce variants of c-myb with qualitatively distinct transcriptional activities that may be involved in transformation and leukemogenesis. Here, by performing a detailed, single molecule assay we found that c-myb alternative RNA splicing was elevated and much more complex in leukemia samples than in cell lines or CD34+ hematopoietic progenitor cells from normal donors. The results revealed that leukemia samples express more than 60 different c-myb splice variants, most of which have multiple alternative splicing events and were not detectable by conventional microarray or PCR approaches. For example, the single molecule assay detected 21 and 22 splice variants containing the 9B and 9S exons, respectively, most of which encoded unexpected variant forms of c-Myb protein. Furthermore, the detailed analysis identified some splice variants whose expression correlated with poor survival in a small cohort of precursor B-ALL samples. Our findings indicate that single molecule assays can reveal complexities in c-myb alternative splicing that have potential as novel biomarkers and could help explain the role of c-Myb variants in the development of human leukemia. The c-Myb transcription factor, a key regulator of proliferation and differentiation in hematopoietic and other cell types, has an N-terminal DNA binding domain and a large C-terminal domain responsible for transcriptional activation, negative regulation and determining target gene specificity. Overexpression and rearrangement of the c- myb gene (MYB) has been reported in some patients with leukemias and other types of cancers, implicating activated alleles of c- myb in the development of human tumors. Alternative RNA splicing can produce variants of c- myb with qualitatively distinct transcriptional activities that may be involved in transformation and leukemogenesis. Here, by performing a detailed, single molecule assay we found that c- myb alternative RNA splicing was elevated and much more complex in leukemia samples than in cell lines or CD34+ hematopoietic progenitor cells from normal donors. The results revealed that leukemia samples express more than 60 different c- myb splice variants, most of which have multiple alternative splicing events and were not detectable by conventional microarray or PCR approaches. For example, the single molecule assay detected 21 and 22 splice variants containing the 9B and 9S exons, respectively, most of which encoded unexpected variant forms of c-Myb protein. Furthermore, the detailed analysis identified some splice variants whose expression correlated with poor survival in a small cohort of precursor B-ALL samples. Our findings indicate that single molecule assays can reveal complexities in c- myb alternative splicing that have potential as novel biomarkers and could help explain the role of c-Myb variants in the development of human leukemia. |
Audience | Academic |
Author | Edwards, Jeremy S Ness, Scott A Zhou, Ye E O'Rourke, John P |
AuthorAffiliation | Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, United States of America University of Florida, United States of America |
AuthorAffiliation_xml | – name: University of Florida, United States of America – name: Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, United States of America |
Author_xml | – sequence: 1 givenname: Ye E surname: Zhou fullname: Zhou, Ye E organization: Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, United States of America – sequence: 2 givenname: John P surname: O'Rourke fullname: O'Rourke, John P – sequence: 3 givenname: Jeremy S surname: Edwards fullname: Edwards, Jeremy S – sequence: 4 givenname: Scott A surname: Ness fullname: Ness, Scott A |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/21853052$$D View this record in MEDLINE/PubMed |
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Copyright | COPYRIGHT 2011 Public Library of Science 2011 Zhou et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. Zhou et al. 2011 |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceived and designed the experiments: SAN. Performed the experiments: YEZ JPO. Analyzed the data: YEZ JSE SAN. Contributed reagents/materials/analysis tools: JPO JSE. Wrote the paper: YEZ SAN. |
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SubjectTerms | Alternative splicing Alternative Splicing - genetics Analysis Assaying Bioindicators Biology Biomarkers Biomarkers, Tumor - genetics c-Myb protein CD34 antigen Cell Line, Tumor Cells (biology) Child Clinical medicine Deoxyribonucleic acid Development and progression DNA DNA microarrays Exons Exons - genetics Gene expression Gene rearrangement Gene regulation Genes Genetic transformation Genomics Health sciences Hematopoietic stem cells Humans Leukemia Leukemogenesis Medicine Mutation MYB gene MYB protein Pediatrics Polymerase Chain Reaction - methods Precursor B-Cell Lymphoblastic Leukemia-Lymphoma - classification Precursor B-Cell Lymphoblastic Leukemia-Lymphoma - genetics Precursors Progenitor cells Proteins Proto-Oncogene Proteins c-myb - genetics Ribonucleic acid RNA RNA, Messenger - genetics RNA, Messenger - metabolism Stem cells Survival Analysis Transcription (Genetics) Transcription activation Transformation Tumor cell lines Tumors |
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Title | Single molecule analysis of c-myb alternative splicing reveals novel classifiers for precursor B-ALL |
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