HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 277, Issue 7, 5256-5264, February 15, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/7/5256    most recent M108895200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lo, H.-R. Articles by Chao, Y.-C. Search for Related Content PubMed PubMed Citation Articles by Lo, H.-R. Articles by Chao, Y.-C. Social Bookmarking                      What's this?

Novel Baculovirus DNA Elements Strongly Stimulate Activities of Exogenous and Endogenous Promoters^*

Huei-Ru Lo, Cheng-Chung Chou, Tzong-Yuan Wu^§, Joyce Pui-Yee Yuen, and Yu-Chan Chao^¶

From the Institute of Molecular Biology, Academia Sinica, Nankang, Taipei 115, Taiwan, Republic of China

A DNA sequence upstream from the polyhedrin gene of baculovirus Autographa californica nucleopolyhedrovirus (AcMNPV) was found to activate strongly the expression of full or minimal promoters derived from AcMNPV and other sources. Promoters tested included the minimal CMV (CMVm) promoter from human cytomegalovirus, the full heat shock 70 promoter from Drosophila, and the minimal p35 promoter from baculovirus. Deletion and mutagenesis analyses showed that this functional polyhedrin upstream (pu) activator sequence contains three open reading frames (ORFs), ORF4, ORF5, and lef2. In plasmid transfection assays, the pu sequence was able to confer high level luciferase expression driven by all of these full or minimal promoters in insect Sf21 cells. A known baculovirus enhancer, the homologous region (hr) of AcMNPV, further enhanced the expression of these promoters. Experiments showed that although multiple hr sequences function in an additive manner, pu and hr together function synergistically, resulting in as much as 18,000-fold promoter activation. Furthermore, a modified CMVm promoter containing pu and/or hr was inserted into the baculovirus genome to drive the luciferase coding region. The CMVm promoter expressed luciferase much earlier, and although it expressed a bit less than did the p10 promoter, the CMVm promoter gave rise to greater luciferase activity. Therefore, we have uncovered a cryptic viral sequence capable of activating a diverse group of promoters. Finally, these experiments demonstrate that synthetic sequences containing pu, hr, and different full or minimal promoters can generate a set of essentially unlimited novel promoters for weak to very strong expression of foreign proteins using baculovirus.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				C. Y.-Y. Liu, C.-H. Wang, W.-K. Hsiao, H.-R. Lo, C. P. Wu, and Y. C. Chao RING and Coiled-Coil Domains of Baculovirus IE2 Are Critical in Strong Activation of the Cytomegalovirus Major Immediate-Early Promoter in Mammalian Cells J. Virol., April 15, 2009; 83(8): 3604 - 3616. [Abstract] [Full Text] [PDF]

				C. Y. Y. Liu, C. H. Wang, J. C. Wang, and Y. C. Chao Stimulation of baculovirus transcriptome expression in mammalian cells by baculoviral transcriptional activators J. Gen. Virol., August 1, 2007; 88(8): 2176 - 2184. [Abstract] [Full Text] [PDF]

				P. Viswanathan, B. Venkaiah, M. S. Kumar, S. Rasheedi, S. Vrati, M. D. Bashyam, and S. E. Hasnain The Homologous Region Sequence (hr1) of Autographa californica Multinucleocapsid Polyhedrosis Virus Can Enhance Transcription from Non-baculoviral Promoters in Mammalian Cells J. Biol. Chem., December 26, 2003; 278(52): 52564 - 52571. [Abstract] [Full Text] [PDF]