The heptanucleotide motif GAGACGC is a key component of a cis-acting promoter element that is critical for SnSAG1 expression in Sarcocystis neurona

Rajshekhar Y. Gaji; Daniel K. Howe

doi:10.1016/j.molbiopara.2009.02.005

The heptanucleotide motif GAGACGC is a key component of a cis-acting promoter element that is critical for SnSAG1 expression in Sarcocystis neurona

Rajshekhar Y. Gaji, Daniel K. Howe

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

The apicomplexan parasite Sarcocystis neurona undergoes a complex process of intracellular development, during which many genes are temporally regulated. The described study was undertaken to begin identifying the basic promoter elements that control gene expression in S. neurona. Sequence analysis of the 5′-flanking region of five S. neurona genes revealed a conserved heptanucleotide motif GAGACGC that is similar to the WGAGACG motif described upstream of multiple genes in Toxoplasma gondii. The promoter region for the major surface antigen gene SnSAG1, which contains three heptanucleotide motifs within 135 bases of the transcription start site, was dissected by functional analysis using a dual luciferase reporter assay. These analyses revealed that a minimal promoter fragment containing all three motifs was sufficient to drive reporter molecule expression, with the presence and orientation of the 5′-most heptanucleotide motif being absolutely critical for promoter function. Further studies should help to identify additional sequence elements important for promoter function and for controlling gene expression during intracellular development by this apicomplexan pathogen.

Original language	English
Pages (from-to)	85-88
Number of pages	4
Journal	Molecular and Biochemical Parasitology
Volume	166
Issue number	1
DOIs	https://doi.org/10.1016/j.molbiopara.2009.02.005
State	Published - Jul 2009

Bibliographical note

Funding Information:
We thank Dr. C.S. Kaetzel for providing plasmids pRL-CMV and pGL2-Basic, and Dr. Vern Carruthers and Dr. Ling Yuan for critical review of the manuscript. We also gratefully acknowledge the technical assistance of Dr. Anthony Sinai and Dr. Robert Molestina. This research was supported by grants from the Amerman Family Foundation and Fort Dodge Animal Health to DKH. Raj Gaji was supported by a Paul Mellon Graduate Student Fellowship in Equine Veterinary Science. Published as Kentucky Agricultural Experiment Station Article No. 06-14-126.

Funding

We thank Dr. C.S. Kaetzel for providing plasmids pRL-CMV and pGL2-Basic, and Dr. Vern Carruthers and Dr. Ling Yuan for critical review of the manuscript. We also gratefully acknowledge the technical assistance of Dr. Anthony Sinai and Dr. Robert Molestina. This research was supported by grants from the Amerman Family Foundation and Fort Dodge Animal Health to DKH. Raj Gaji was supported by a Paul Mellon Graduate Student Fellowship in Equine Veterinary Science. Published as Kentucky Agricultural Experiment Station Article No. 06-14-126.

Funders	Funder number
Fort Dodge Animal Health
Amerman Family Foundation

Keywords

Apicomplexa
Coccidia
Gene regulation
Luciferase
Promoter
Sarcocystis neurona
Transcription
Transfection

ASJC Scopus subject areas

Parasitology
Molecular Biology

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10.1016/j.molbiopara.2009.02.005

Cite this

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title = "The heptanucleotide motif GAGACGC is a key component of a cis-acting promoter element that is critical for SnSAG1 expression in Sarcocystis neurona",

abstract = "The apicomplexan parasite Sarcocystis neurona undergoes a complex process of intracellular development, during which many genes are temporally regulated. The described study was undertaken to begin identifying the basic promoter elements that control gene expression in S. neurona. Sequence analysis of the 5′-flanking region of five S. neurona genes revealed a conserved heptanucleotide motif GAGACGC that is similar to the WGAGACG motif described upstream of multiple genes in Toxoplasma gondii. The promoter region for the major surface antigen gene SnSAG1, which contains three heptanucleotide motifs within 135 bases of the transcription start site, was dissected by functional analysis using a dual luciferase reporter assay. These analyses revealed that a minimal promoter fragment containing all three motifs was sufficient to drive reporter molecule expression, with the presence and orientation of the 5′-most heptanucleotide motif being absolutely critical for promoter function. Further studies should help to identify additional sequence elements important for promoter function and for controlling gene expression during intracellular development by this apicomplexan pathogen.",

keywords = "Apicomplexa, Coccidia, Gene regulation, Luciferase, Promoter, Sarcocystis neurona, Transcription, Transfection",

author = "Gaji, {Rajshekhar Y.} and Howe, {Daniel K.}",

note = "Funding Information: We thank Dr. C.S. Kaetzel for providing plasmids pRL-CMV and pGL2-Basic, and Dr. Vern Carruthers and Dr. Ling Yuan for critical review of the manuscript. We also gratefully acknowledge the technical assistance of Dr. Anthony Sinai and Dr. Robert Molestina. This research was supported by grants from the Amerman Family Foundation and Fort Dodge Animal Health to DKH. Raj Gaji was supported by a Paul Mellon Graduate Student Fellowship in Equine Veterinary Science. Published as Kentucky Agricultural Experiment Station Article No. 06-14-126. ",

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N2 - The apicomplexan parasite Sarcocystis neurona undergoes a complex process of intracellular development, during which many genes are temporally regulated. The described study was undertaken to begin identifying the basic promoter elements that control gene expression in S. neurona. Sequence analysis of the 5′-flanking region of five S. neurona genes revealed a conserved heptanucleotide motif GAGACGC that is similar to the WGAGACG motif described upstream of multiple genes in Toxoplasma gondii. The promoter region for the major surface antigen gene SnSAG1, which contains three heptanucleotide motifs within 135 bases of the transcription start site, was dissected by functional analysis using a dual luciferase reporter assay. These analyses revealed that a minimal promoter fragment containing all three motifs was sufficient to drive reporter molecule expression, with the presence and orientation of the 5′-most heptanucleotide motif being absolutely critical for promoter function. Further studies should help to identify additional sequence elements important for promoter function and for controlling gene expression during intracellular development by this apicomplexan pathogen.

AB - The apicomplexan parasite Sarcocystis neurona undergoes a complex process of intracellular development, during which many genes are temporally regulated. The described study was undertaken to begin identifying the basic promoter elements that control gene expression in S. neurona. Sequence analysis of the 5′-flanking region of five S. neurona genes revealed a conserved heptanucleotide motif GAGACGC that is similar to the WGAGACG motif described upstream of multiple genes in Toxoplasma gondii. The promoter region for the major surface antigen gene SnSAG1, which contains three heptanucleotide motifs within 135 bases of the transcription start site, was dissected by functional analysis using a dual luciferase reporter assay. These analyses revealed that a minimal promoter fragment containing all three motifs was sufficient to drive reporter molecule expression, with the presence and orientation of the 5′-most heptanucleotide motif being absolutely critical for promoter function. Further studies should help to identify additional sequence elements important for promoter function and for controlling gene expression during intracellular development by this apicomplexan pathogen.

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The heptanucleotide motif GAGACGC is a key component of a cis-acting promoter element that is critical for SnSAG1 expression in Sarcocystis neurona

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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