This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wahlers, A.
Right arrow Articles by Baum, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wahlers, A.
Right arrow Articles by Baum, C.

 Previous Article  |  Next Article 

Journal of Virology, January 2002, p. 303-312, Vol. 76, No. 1
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.1.303-312.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

In Vivo Analysis of Retroviral Enhancer Mutations in Hematopoietic Cells: SP1/EGR1 and ETS/GATA Motifs Contribute to Long Terminal Repeat Specificity

Anke Wahlers,1 Peter F. Zipfel,2 Maike Schwieger,1 Wolfram Ostertag,1 and Christopher Baum3*

Department of Cell and Virus Genetics, Heinrich Pette Institute, D-20251 Hamburg,1 Hans Knoell Institute for Natural Products Research, D-07745 Jena,2 Department of Hematology and Oncology, Medical School Hannover, D-30625 Hannover, Germany3

Received 9 May 2001/ Accepted 26 September 2001

The objective of this work was to identify, in the context of chromosomally integrated DNA, the contribution of defined transcription factor binding motifs to the function of a complex retrovirus enhancer in hematopoietic cells in vivo. Repopulating murine hematopoietic cells were transduced with equal gene dosages of replication-incompetent retrovirus vectors encoding enhanced green fluorescent protein. Enhancer sequences were derived from mouse spleen focus-forming virus. Destruction of GC-rich sites representing overlapping targets for SP1 or EGR1 uniformly attenuated gene expression (~25 to 70% of wild-type levels) in all hematopoietic lineages, as shown by multicolor flow cytometry of peripheral blood and bone marrow cells at various time points posttransplantation. In contrast, a point mutation within a dual ETS/GATA motif that abolished transactivation by ETS factors but not by GATA-1 slightly increased activity in erythroid cells and significantly attenuated enhancer function in T lymphocytes. This study shows that controlled gene transfer in transplantable hematopoietic cells allows a functional analysis of distinct cis elements within a complex retrovirus enhancer, as required for the characterization and engineering of various cellular and viral regulatory sequences in basic research and gene therapy.

* Corresponding author. Mailing address: Experimental Cell Therapy, Dept. of Hematology and Oncology, Medical School, OE 5120, Carl-Neuberg-Str. 1, D-30625 Hannover, Germany. Phone: 49-511-532-4525. Fax: 49-511-532-8525. E-mail: baum.christopher{at}mh-hannover.de.

Journal of Virology, January 2002, p. 303-312, Vol. 76, No. 1
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.1.303-312.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Cammenga, J., Niebuhr, B., Horn, S., Bergholz, U., Putz, G., Buchholz, F., Lohler, J., Stocking, C. (2007). RUNX1 DNA-Binding Mutants, Associated with Minimally Differentiated Acute Myelogenous Leukemia, Disrupt Myeloid Differentiation. Cancer Res. 67: 537-545 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»