A cis-regulatory element within the 5' flanking region of arylsulfatase gene of sea urchin, Hemicentrotus pulcherrimus.
The 5' flanking region of the sea urchin Hemicentrotus pulcherrimus arylsulfatase (Ars) gene was scanned to define cis-regulatory elements required for proper expression congruent to that of the endogenous gene. The region between -100 bp and +38 bp from the transcription start site contains minimum information required for temporal initiation of transcription of the Ars gene. Progressive deletion analysis of Ars-luciferase reporter constructs containing the Ars sequence from -3484 bp to +38 bp suggests the existence of several cis-regulatory elements within this region. Results from luciferase assays of internal deletion mutants show strong enhancer activity detected within the sequence from -194 bp to -144 bp. By gel mobility shift assay, we have identified a nuclear factor that interacts sequence-specifically with this 50 bp region, and appears in a developmental stage-specific manner. Further deletion analysis determined that the enhancer activity lies within a 22 bp sequence between -186 bp and -164 bp.
Morokuma
J
J
Graduate Department of Gene Science, Faculty of Science, Hiroshima University, Japan.
Akasaka
K
K
Mitsunaga-Nakatsubo
K
K
Shimada
H
H
D000818:Animals; D019677:Artificial Gene Fusion; D001192:Arylsulfatases / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004742:Enhancer Elements, Genetic; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005809:Genes, Regulator; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D009865:Oocytes / Q000201:enzymology; D012617:Sea Urchins / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D017384:Sequence Deletion
D017931:DNA Primers; D004247:DNA; D008156:Luciferases; D001192:Arylsulfatases