Tissue-specific expression of a Ca(2+)-activated K+ channel is controlled by multiple upstream regulatory elements.
The electrical properties of a cell are produced by the complement of ion channels that it expresses. To understand how ion-channel gene expression is regulated, we are studying the tissue-specific regulation of the slowpoke (slo) Ca(2+)-activated K+ channel gene. This gene is expressed in the central and peripheral nervous system, in midgut and tracheal cells, and in the musculature of Drosophila melanogaster. The entire transcriptional control region has been cloned previously and shown to reproduce the tissue and developmental expression pattern of the endogenous gene. Here we demonstrate that s/o has at least four promoters distributed over approximately 4.5 kb of DNA. Promoter C1 and C1c display a TATA box-like sequence at the appropriate distance from the transcription start site. Promoters C1b and C2, however, are TATA-less promoters. C1, C1b, and C1c transcripts differ in their leader sequence but share a common translation start site. C2 transcripts incorporate a new translation start site that appends 17 amino acids to the N terminus of the encoded protein. Deletion analysis was used to identify sequences important for tissue-specific expression. We used a transgenic in vivo expression system in which all tissues and developmental stages can be assayed easily. Six nested deletions were transformed into Drosophila, and the expression pattern was determined using a lacZ reporter in both dissected tissues and sectioned animals. We have identified different sequences required for expression in the CNS, midgut, tracheal cells, and muscle.
Brenner
R
R
Department of Zoology, University of Texas at Austin 78712-1064, USA.
Thomas
T O
TO
Becker
M N
MN
Atkinson
N S
NS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000502:physiology*; D002490:Central Nervous System / Q000502:physiology; D004331:Drosophila melanogaster; D017353:Gene Deletion; D015870:Gene Expression; D005809:Genes, Regulator; D007422:Intestines / Q000502:physiology; D008969:Molecular Sequence Data; D009132:Muscles / Q000502:physiology; D015221:Potassium Channels / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014132:Trachea / Q000502:physiology; D014158:Transcription, Genetic
D015221:Potassium Channels; D002118:Calcium