Nucleic Acids Research, 1994, Vol. 22, No. 23 5099-5111
© 1994
MOLECULAR BIOLOGY |
Identification and characterization of multiple A/T-rich cisacting elements that control expression from Dictyostelium actin promoters: the Dictyostelium actin upstream activating sequence confers growth phase expression and has enhancer-like properties
Department of Biology, Center for Molecular Genetics, University of Califomia–San Diego 9500 Gilman Drive, La Jolla, CA 92093-0634, USA
*To whom correspondence shoul be addressed
Received May 6, 1994. Revised October 14, 1994. Accepted October 14, 1994.
The promoter elements in the Dictyostelium actin 15 and actin 6 genes required for full growth phase expression were identified by assaying promoter/ luciferase reporter constructs. We find that these promoters contain common c/s-acting elements, an actin upstream activating sequence (UAS) and sequences proximal to the transcription start site that overlap with a poly(dT) region. The actin 15 promoter has two additional c/s-acting elements not present in the actin 6 promoter that may account for the higher level of expression from the actin 15 promoter. All of the identified promoter elements are unusual for Dictyostelium in that they are all A/T-rich. Two c/sacting elements, the actin UAS and the poly(dT) domain were studied in greater detail. The actin UAS was tested on a heterologous promoter from the prespore-specific gene SP60 and shown to have the ability to confer growth phase expression. The actin UAS also exhibited the ability to function in a distance- and orientationindependent manner and activate expression synergistically when present in two copies. The poly(dT) domain of the actin 15 promoter was studied in greater detail by using a genetic selection scheme to define parameters that effect the strength of this element. This element is comprised of 45 consecutive dT residues immediately upstream of the putative TATA box. We show that the length of the homopolymer dT region correlates with the expression level of the promoter. The poly(dT) element is also shown to function to promote wild-type levels of expression with smallm deviations in the sequence, indicating that the element is not required to be homopolymeric to function.
+Present address: Department of Biological Chemistry, Center for Health Sciences, UCLA school of Medicine, 10833 Leconte Avenue, Los angeles, CA 90024-1737, USA
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