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 Mesner, L. D.
Right arrow Articles by Hamlin, J. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mesner, L. D.
Right arrow Articles by Hamlin, J. L.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, February 2003, p. 804-814, Vol. 23, No. 3
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.3.804-814.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Dihydrofolate Reductase Origin of Replication Does Not Contain Any Nonredundant Genetic Elements Required for Origin Activity

L. D. Mesner, X. Li, P. A. Dijkwel, and J. L. Hamlin*

Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, Virginia 22908

Received 19 August 2002/ Returned for modification 24 September 2002/ Accepted 1 November 2002

The Chinese hamster dihydrofolate reductase (DHFR) origin of replication consists of a broad zone of potential initiation sites scattered throughout a 55-kb intergenic spacer, with at least three sites being preferred (ori-ß, ori-ß', and ori-{gamma}). We previously showed that deletion of the most active site or region (ori-ß) has no demonstrable effect on initiation in the remainder of the intergenic spacer nor on the time of replication of the DHFR locus as a whole. In the present study, we have now deleted ori-ß', both ori-ß and ori-ß', an 11-kb region just downstream from the DHFR gene, or the central ~40-kb core of the spacer. The latter two deletions together encompass >95% of the initiation sites that are normally used in this locus. Two-dimensional gel analysis shows that initiation still occurs in the early S phase in the remainder of the intergenic spacer in each of these deletion variants. Even removal of the 40-kb core fails to elicit a significant effect on the time of replication of the DHFR locus in the S period; indeed, in the truncated spacer that remains, the efficiency of initiation actually appears to increase relative to the corresponding region in the wild-type locus. Thus, if replicators control the positions of nascent strand start sites in this complex origin, either (i) there must be a very large number of redundant elements in the spacer, each of which regulates initiation only in its immediate environment, or (ii) they must lie outside the central core in which the vast majority of nascent strand starts occur.


* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA 22908. Phone: (434) 924-5858. Fax: (434) 924-1789. E-mail: jlh2d{at}virginia.edu.


Molecular and Cellular Biology, February 2003, p. 804-814, Vol. 23, No. 3
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.3.804-814.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.




This article has been cited by other articles:

  • Houchens, C. R., Lu, W., Chuang, R.-Y., Frattini, M. G., Fuller, A., Simancek, P., Kelly, T. J. (2008). Multiple Mechanisms Contribute to Schizosaccharomyces pombe Origin Recognition Complex-DNA Interactions. J. Biol. Chem. 283: 30216-30224 [Abstract] [Full Text]  
  • Hayashida, T., Oda, M., Ohsawa, K., Yamaguchi, A., Hosozawa, T., Locksley, R. M., Giacca, M., Masai, H., Miyatake, S. (2006). Replication Initiation from a Novel Origin Identified in the Th2 Cytokine Cluster Locus Requires a Distant Conserved Noncoding Sequence. J. Immunol. 176: 5446-5454 [Abstract] [Full Text]  
  • Sasaki, T., Ramanathan, S., Okuno, Y., Kumagai, C., Shaikh, S. S., Gilbert, D. M. (2006). The Chinese Hamster Dihydrofolate Reductase Replication Origin Decision Point Follows Activation of Transcription and Suppresses Initiation of Replication within Transcription Units. Mol. Cell. Biol. 26: 1051-1062 [Abstract] [Full Text]  
  • Wang, J., Lindner, S. E., Leight, E. R., Sugden, B. (2006). Essential Elements of a Licensed, Mammalian Plasmid Origin of DNA Synthesis. Mol. Cell. Biol. 26: 1124-1134 [Abstract] [Full Text]  
  • Patel, P. K., Arcangioli, B., Baker, S. P., Bensimon, A., Rhind, N. (2006). DNA Replication Origins Fire Stochastically in Fission Yeast. Mol. Biol. Cell 17: 308-316 [Abstract] [Full Text]  
  • Jeon, Y., Bekiranov, S., Karnani, N., Kapranov, P., Ghosh, S., MacAlpine, D., Lee, C., Hwang, D. S., Gingeras, T. R., Dutta, A. (2005). Temporal profile of replication of human chromosomes. Proc. Natl. Acad. Sci. USA 102: 6419-6424 [Abstract] [Full Text]  
  • Mesner, L. D., Hamlin, J. L. (2005). Specific signals at the 3' end of the DHFR gene define one boundary of the downstream origin of replication. Genes Dev. 19: 1053-1066 [Abstract] [Full Text]  
  • Casper, J. M., Kemp, M. G., Ghosh, M., Randall, G. M., Vaillant, A., Leffak, M. (2005). The c-myc DNA-unwinding Element-binding Protein Modulates the Assembly of DNA Replication Complexes in Vitro. J. Biol. Chem. 280: 13071-13083 [Abstract] [Full Text]  
  • Kemp, M. G., Ghosh, M., Liu, G., Leffak, M. (2005). The histone deacetylase inhibitor trichostatin A alters the pattern of DNA replication origin activity in human cells. Nucleic Acids Res 33: 325-336 [Abstract] [Full Text]  
  • Chang, V. K., Donato, J. J., Chan, C. S., Tye, B. K. (2004). Mcm1 Promotes Replication Initiation by Binding Specific Elements at Replication Origins. Mol. Cell. Biol. 24: 6514-6524 [Abstract] [Full Text]  
  • Altman, A. L., Fanning, E. (2004). Defined Sequence Modules and an Architectural Element Cooperate To Promote Initiation at an Ectopic Mammalian Chromosomal Replication Origin. Mol. Cell. Biol. 24: 4138-4150 [Abstract] [Full Text]  
  • Wang, L., Lin, C.-M., Brooks, S., Cimbora, D., Groudine, M., Aladjem, M. I. (2004). The Human {beta}-Globin Replication Initiation Region Consists of Two Modular Independent Replicators. Mol. Cell. Biol. 24: 3373-3386 [Abstract] [Full Text]  
  • Saha, S., Shan, Y., Mesner, L. D., Hamlin, J. L. (2004). The promoter of the Chinese hamster ovary dihydrofolate reductase gene regulates the activity of the local origin and helps define its boundaries. Genes Dev. 18: 397-410 [Abstract] [Full Text]  
  • Bowmaker, M., Yang, M. Y., Yasukawa, T., Reyes, A., Jacobs, H. T., Huberman, J. A., Holt, I. J. (2003). Mammalian Mitochondrial DNA Replicates Bidirectionally from an Initiation Zone. J. Biol. Chem. 278: 50961-50969 [Abstract] [Full Text]