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J. Biol. Chem., Vol. 275, Issue 44, 34597-34608, November 3, 2000
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From the Department of Genetics, Boyer Center for Molecular
Medicine, Yale University School of Medicine,
New Haven, Connecticut 06510
The gene for the human leukocyte-specific
transcript 1 (LST1) encodes a small protein that modulates immune
responses and cellular morphogenesis. The LST1 transcripts are
expressed at high levels in dendritic cells. Because of the complex
splicing pattern, use of alternative 5'-untranslated exons, and
a biologically interesting pattern of expression of LST1 mRNA, we
studied the human LST1 gene promoter and regulatory elements. We
identified an additional upstream 5'-untranslated exon in U937
monocytic cells. Transient transfection studies demonstrated that the
combination of regions from
1363 to
621 with
112 to
54,
relative to the translation start codon, produced the highest level of
transcripts from among the various constructs tested, but the pattern
of transcripts produced was only a subset of those produced from the
endogenous gene. DNase I footprinting analysis and electrophoretic
mobility shift assays showed that oligonucleotide probes corresponding to three regions,
1171 to
1142 (BI),
1136 to
1111 (BII), and
783 to
751 (BIV), bound proteins in U937 nuclear extracts.
Competition and supershift electrophoretic mobility shift assay did not
identify any known transcription factors responsible for BII probe
binding. These studies suggest that a novel DNA-binding site and
interaction of multiple regulatory elements may be involved in
mediating the expression of the various forms of LST1 mRNA.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AW928284, AW925228, AW927279, and AW927280.
To whom correspondence should addressed: Dept. of Genetics, Boyer
Center for Molecular Medicine, Yale University School of Medicine, 295 Congress Ave., New Haven, CT 06510. Tel.: 203-737-2282; Fax:
203-737-2286; E-mail: Sherman.Weissman@yale.edu.
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