Structure and Function of the MAC-1 Gene

Roberts, Thomas M.

Abstract

The objective of the project is to obtain full length genomic DNA clones and cDNA clones for both the alpha and beta-chains of the functionally important cell surface antigen, MAC-1, and to characterize these clones in detail. The complete DNA sequence of the cDNA clones will be determined and from this the protein sequence of MAC-1 deduced. Extensive studies on the relationship between structure and function of the protein will be undertaken using a variety of mutagenesis protocols. Mutants in the protein will be assayed for function using a very efficient protocol based upon an easily rescued retrovirus vector. The promoters for both MAC-1 genes will be mapped on the genomic clones and further characterized by genetic analysis. Changes in the level of methylation of the gene and of the DNAse reactivity of the gene's chromatin will be studied as a function of macrophage differentiation. Proteins which bind to the genes control region will be characterized by Crothers' gel analysis.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Allergy and Infectious Diseases (NIAID)
• Type: Research Project (R01)
• Project #: 2R01AI020316-04A1
• Application #: 3129879
• Study Section: Experimental Immunology Study Section (EI)
• Project Start: 1983-08-01
• Project End: 1990-03-31
• Budget Start: 1987-04-01
• Budget End: 1988-03-31
• Support Year: 4
• Fiscal Year: 1987

Institution

• Name: Dana-Farber Cancer Institute
• DUNS #: 149617367
• City: Boston
• State: MA
• Country: United States
• Zip Code: 02115

Publications

Corbi, A L; Garcia-Aguilar, J; Springer, T A (1990) Genomic structure of an integrin alpha subunit, the leukocyte p150,95 molecule. J Biol Chem 265:2782-8

Corbi, A L; Kishimoto, T K; Miller, L J et al. (1988) The human leukocyte adhesion glycoprotein Mac-1 (complement receptor type 3, CD11b) alpha subunit. Cloning, primary structure, and relation to the integrins, von Willebrand factor and factor B. J Biol Chem 263:12403-11

Kishimoto, T K; O'Connor, K; Lee, A et al. (1987) Cloning of the beta subunit of the leukocyte adhesion proteins: homology to an extracellular matrix receptor defines a novel supergene family. Cell 48:681-90

Gee, C E; Griffin, J; Sastre, L et al. (1986) Differentiation of myeloid cells is accompanied by increased levels of pp60c-src protein and kinase activity. Proc Natl Acad Sci U S A 83:5131-5

Sastre, L; Roman, J M; Teplow, D B et al. (1986) A partial genomic DNA clone for the alpha subunit of the mouse complement receptor type 3 and cellular adhesion molecule Mac-1. Proc Natl Acad Sci U S A 83:5644-8

Sastre, L; Kishimoto, T K; Gee, C et al. (1986) The mouse leukocyte adhesion proteins Mac-1 and LFA-1: studies on mRNA translation and protein glycosylation with emphasis on Mac-1. J Immunol 137:1060-5