Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor

Y. Lee, S. E. Stepp, P. R. Kumaresan, P. A. Mathew

Research output: Contribution to journalArticle

Abstract

We have previously identified, cloned and characterized a receptor, 2B4, expressed on murine NK cells and a subset of T cells. Functional studies indicate that ligation of 2B4 transduce activation signals and is associated with increased target cell lysis. 2B4 is a member of the immunoglobulin superfamily and shows homology to CD48, LFA-3 and SLAM. In order to further understand the transcriptional regulation of the 2B4 gene, the promoter region was subcloned and analyzed. Using nuclear protein extract from a T cell line, CTLL-2, the interaction of the promoter with nuclear proteins was studied by electrophoretic mobility shift assay (EMSA).The result showed that a 100bp 5′ fragment (-89 to +11) of the promoter region interacted with nuclear protein(s) expressed only in CTLL-2 cells, which express 2B4, whereas, such specific binding was not seen in Jurkat, Peer (both T cell line ) or Sp2/o (B cell line ) cells which do not express 2B4. Competition EMSA showed that the binding was sequence specific. Nested deletion of the 2B4 promoter region and analysis by the reporter gene chloramphenicol acetyl transferase yielded varying levels of CAT expression suggesting the presence of two cis-acting elements in the 2B4 promoter.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number5
StatePublished - Mar 20 1998
Externally publishedYes

Fingerprint

Natural Killer Cell Receptors
T-cells
Cloning
natural killer cells
Nuclear Proteins
Genetic Promoter Regions
Organism Cloning
molecular cloning
Electrophoretic mobility
promoter regions
Electrophoretic Mobility Shift Assay
nuclear proteins
Cell Line
receptors
Assays
mice
chloramphenicol acetyltransferase
Genes
CD58 Antigens
T-lymphocytes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Lee, Y., Stepp, S. E., Kumaresan, P. R., & Mathew, P. A. (1998). Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor. FASEB Journal, 12(5).

Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor. / Lee, Y.; Stepp, S. E.; Kumaresan, P. R.; Mathew, P. A.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

Lee, Y, Stepp, SE, Kumaresan, PR & Mathew, PA 1998, 'Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor', FASEB Journal, vol. 12, no. 5.
Lee Y, Stepp SE, Kumaresan PR, Mathew PA. Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor. FASEB Journal. 1998 Mar 20;12(5).
Lee, Y. ; Stepp, S. E. ; Kumaresan, P. R. ; Mathew, P. A. / Cloning and analysis of the promoter region of 2B4, a murine NK cell receptor. In: FASEB Journal. 1998 ; Vol. 12, No. 5.
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