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Nucleic Acids Research, 1990, Vol. 18, No. 14 4185-4190
© 1990

GENOME STRUCTURE AND MAPPING

Comparison of the promoter regions of H-2Kb and H-2Kbm1 class I MHC genes

Kazuo Ozawa1, Fujiko Saka1, Issay Kitabayashi1, Takashi lmai1, Eiichi Soeda1, Alain Israel2, Gabriel Gachelin2,3 and Kazushige Yokoyama1,3,*

1Gene Bank, Life Science Tsukuba Research Center, RIKEN (The Institute of Physical and Chemical Research), Koyadai 3-1-1 Tsukuba, Ibaraki 305, Japan 2Department of Immunology, Institut Pasteur 25 rue du Dr Roux, 75724 Paris Cedex 15, France 3Frontier Research Program, Life Science Tsukuba Research Center, RIKEN (The Institute of Physical and Chemical Research), Koyadai 3-1-1 Tsukuba, Ibaraki 305, Japan

* To whom correspondence should be addressed

Received March 29, 1990. Revised May 14, 1990. Accepted May 14, 1990.

The 2.0 kb-long nucleotide sequences of the promoter regions of two closely related class I genes of the mouse major histocompatibility complex (H-2Kb and H-2Kbm1) have been determined and compared. The promoter sequence of the H-2Kbm1 gene differs from that of the H-2Kb gene by a single deletion of a ‘C’ at position –456 in the upstream region of H-2Kbm1 gene. The actual existence of this deletion of a single base in genomic DNA has been verified by genomic DNA hybridization, using oligonucleotide probes specific for H-2Kbm1 or H-2Kb respectively. The effect on the enhancer activity of H-2Kbm1 promoter region of the difference at position –456 has been analyzed by the chloramphenicol acetyltransferase (CAT) assay, using appropriate Ddel fragments (–533 to –408 for H-2Kbm1; – 534 to – 408 for H-2Kb) cloned downstream of pH-2(367)CAT gene construct. The CAT activity determined by the H-2Kbm1 fragment was about 3-fold higher than that of H-2Kb, a result which probably accounts for the higher level of the H-2Kbm1 transcript and antigen in lymph node cells.


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