An intronic silencer of the mouse perforin gene.
We have previously shown that the perforin gene locus is comprised of eight DNase I hypersensitive sites (DHS). Seven (DHS I-DHS VII) of them were CTL-specific whereas one (DHS VIII) in the second intron was expressed in a wide range of cell types. DHS VIII was highly AT-rich (75%) and was comprised of multiple sets of high mobility group (HMG)-I/Y binding site, two potential Special AT-rich Binding protein (SATB-1)-binding sites, and a long stretch of CTAT repeats, indicating that DHS VIII may relate to nuclear matrix-associated region (MAR). When DHS VIII was inserted into the perforin promoter-driven luciferase gene, it silenced the reporter gene transcription in CTLL-R8 cells in an orientation- and distance-independent manner. Moreover, this silencing effect was also observed in other promoters in a variety of non-CTL cell lines, suggesting that DHS VIII exerted a global silencing effect. Deletion analysis and gel-shift assays indicated that the silencing effect was mediated by the CTAT repeats and its binding protein called CTAT repeats-binding protein (CRBP).
Youn
Byung-Soo
BS
The Immunomodulation Research Center and Department of Biological Sciences, University of Ulsan, Korea. bskwon@uou.ulsan.ac.kr
Lim
Chae Lyul
CL
Shin
Man Kyun
MK
Hill
Jams M
JM
Kwon
Byoung S
BS
D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I; D020868:Gene Silencing; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008156:Luciferases / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism; D020080:Tandem Repeat Sequences; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D054353:Perforin; D004247:DNA; D008156:Luciferases; D003850:Deoxyribonuclease I