Isolation and characterization of the rat SND p102 gene promoter: putative role for nuclear factor-Y in regulation of transcription.
In this work, we report the isolation and characterization of a 1,688-bp sequence corresponding to the promoter region of the rat endoplasmic reticulum (ER) cholesterol ester hydrolase gene, renamed as staphylococcal nuclease domain-containing protein of 102 kDa (SND p102) in GenBank database according to the structural properties and molecular weight of the protein. The transcription start site was located 216 bases upstream of the ATG start codon by RNA ligase mediated-rapid amplification of cDNA ends (RLM-RACE). Bioinformatic analysis of the isolated sequence revealed a lack of typical promoter TATA box and the presence of GC-rich motifs and CCAAT boxes recognized by Sp 1 and nuclear factor-Y among other putative binding sites for a number of transcription factors implicated in both basal and regulated processes. Electrophoretic mobility shift and supershift assays using nuclear extracts from human (HepG2) and rat (McA-RH7777) hepatoma cells demonstrated that nuclear factor-Y (NF-Y) transcription factor bound to the core sequences at (-257, -253), (-290, -286), and (-370, -366) upstream translation initiation site. The absence of TATA box and the location and reverse orientation of the CCAAT boxes in the promoter region strongly suggest a role for NF-Y in the regulation of transcription of SND p102 gene.
Rodríguez
Lorena
L
Department of Physiology, Faculty of Medicine and Dentistry, University of the Basque Country, Sarriena s/n, 48940-Leioa, Bizkaia, Spain.
Bartolomé
Nerea
N
Ochoa
Begoña
B
Martínez
María J
MJ
D000818:Animals; D001483:Base Sequence; D023081:CCAAT-Binding Factor / Q000502:physiology*; D045744:Cell Line, Tumor; D004720:Endonucleases; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000502:physiology; D011401:Promoter Regions, Genetic; D051381:Rats; D016385:TATA Box / Q000235:genetics; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic
D023081:CCAAT-Binding Factor; D009687:Nuclear Proteins; D014157:Transcription Factors; C402244:nuclear factor Y; D004720:Endonucleases; C490639:Snd1 protein, rat