Identification of a negative regulatory DNA element for neuronal BC1 RNA expression by RNA polymerase III.
BC1 RNA is a neuronal cell-specific RNA polymerase III (Pol III) transcript. The BC1 RNA gene has plural types of Pol III promoters, in addition to which an E-box sequence (E2 site) acts as a transcriptional activator, which is recognized by a brain-specific protein(s). Using an in vitro transcription system, we found that the upstream region of the BC1 RNA gene contained a sequence that interfered with the activity of the E-box element in a distance-independent manner. A tandem repeat within this sequence, which was weakly homologous with the neuron-restrictive silencer element (NRSE) found in the Pol II system, was recognized by a brain nuclear protein. Consistently, the transcriptional activity increased by deleting the tandem repeat sequence. We called this BC1 RNA-repressing element BCRE. The DNA-binding specificities of BCRE-binding protein differed from that of NRSE-binding protein (NRSF). A similar protein with an ability to bind to BCRE was also found in liver and kidney. Furthermore, the glutamate analog kainic acid increased the DNA-binding of both E2 site-binding protein and BCRE-binding protein, and then the levels of BC1 RNA also increased transiently. Our results suggested that both positive and negative regulatory elements contribute to neuronal BC1 RNA expression.
Kobayashi
S
S
Department of Biochemistry, College of Pharmacy, Nihon University, Narashinodai, Funabashi, 274-8555, Chiba, Japan.
Kamo
S
S
Ohmae
A
A
Agui
K
K
Li
Y
Y
Anzai
K
K
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D015536:Down-Regulation; D005786:Gene Expression Regulation; D007608:Kainic Acid; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010957:Plasmids; D012320:RNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D020733:RNA, Small Cytoplasmic / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D012640:Seizures / Q000139:chemically induced / Q000378:metabolism; D014158:Transcription, Genetic
C117734:BC1 RNA; D004268:DNA-Binding Proteins; D020733:RNA, Small Cytoplasmic; D012320:RNA Polymerase III; D007608:Kainic Acid