Structure of the murine lck gene and its rearrangement in a murine lymphoma cell line.

Garvin, Alex M.; Pawar, Sagar; Marth, Jamey D.; Perlmutter, Roger M.

doi:10.1128/mcb.8.8.3058

Cited by 79 publications

(88 citation statements)

References 27 publications

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“…The distal promoter lies upstream of this untranslated exon, and the proximal promoter is present within the 5Ј-UTR intron. This intron is Ͼ40 kb in humans (22; UCSC Human Genome Browser) and Ͼ10 kb in mouse (17). The 5Ј-RACE analysis of the Fugu lck cDNA from the kidney, gills, and spleen revealed the presence of a 1.4-kb intron in the 5Ј-UTR located 5 bp upstream of the lck initiation codon and mapped the transcription start site to 154 bp upstream of this intron (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The Fugu lck gene consists of 12 coding exons similar to the human and mouse lck genes (17,22). It codes for a protein of 502 aa, which is 71% identical to the human, mouse, and chicken p56 lck (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Consequently the transcripts generated from the proximal promoter are found only in the thymus whereas the distal promoter transcripts are found both in the thymus and peripheral T cells. The proximal and distal promoters of lck are separated by Ͼ40 kb and 10 kb in humans (UCSC Human Genome browser at http:͞͞ genome.ucsc.edu) and mice (17), respectively.…”

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confidence: 99%

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Conserved regulation of the lymphocyte-specific expression oflckin the Fugu and mammals

Brenner

Venkatesh

Yap

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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The lck gene encodes a lymphocyte-specific protein-tyrosine kinase that is implicated in T cell maturation and signaling. In mammals, the transcription of the lck gene is regulated by two independent promoters, the proximal promoter, which is active in thymocytes, and the distal promoter, which dominates in mature T cells. In the human and mouse lck gene loci, the two promoter elements are separated by at least 40 kb and 10 kb, respectively. In this study, we have cloned and sequenced 60 kb from the pufferfish (Fugu rubripes) lck locus. The promoter region of the Fugu lck spans only 4.2 kb and contains a proximal and a distal promoter in the 2.3-kb region adjacent to the coding sequence. By generating transgenic mice, we have demonstrated that the compact promoter of the Fugu lck contains regulatory elements that direct expression to lymphoid organs of mice. We were able to localize the regulatory elements to a short region of 830 bp without losing specificity to cultured human T cell line. These results show that the basic mechanisms that mediate lymphocyte-specific expression are conserved between teleosts and mammals. The short promoter of the Fugu lck isolated by us offers a powerful tool for labeling T cells, targeting expression, and manipulating T cell activity in fishes as well as in mammals.T he specificity of gene expression for cell types or developmental stages and the response of genes to physiological stimuli are mediated through a combinatorial interaction of promoter sequences, enhancers, and suppressors. These regulatory elements are composed of short stretches of DNA that are generally found in the promoter region of genes but also can be located in introns or dispersed over many kilobases upstream and downstream of genes (1). Finding these short elements in mammalian genomes is a formidable task because the intergenic regions in these genomes are large and complex. The genome of the pufferfish, Fugu rubripes, which is approximately eight times smaller than the human and mouse genomes, contains compact intergenic and intronic regions that are devoid of repetitive sequences (2, 3). This genome offers an attractive model for the rapid screening of noncoding sequences for conserved putative regulatory elements. Conserved regulatory elements driving cell-and stage-specific expression of developmental control genes such as the Hoxb-4, Otx2, Wnt-1, and Pax6 were identified by this strategy (4-7). Conserved regulatory elements were identified in the Fugu neurohypophyseal gene that express specifically in a subset of neurons in the hypothalamus of transgenic rats (8), and the Fugu tyrosinase gene was shown to contain conserved elements that directed melanocyte and retinal pigment epithelium-specific expression in transgenic mice (9). The hypothesis can be proposed that, where physiological systems show conservation over long stretches of evolutionary time, the genes specifying these processes are likely to be conserved not only in their coding sequences but also in their regulatory sequences. The immun...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Conserved regulation of the lymphocyte-specific expression oflckin the Fugu and mammals

Brenner

Venkatesh

Yap

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…The complete coding sequence of rat Numb cDNA (nt 196-2007) (23) was cloned into the BamHI site of p1017, which contains the Lck proximal promoter and portions of the human growth hormone gene, including a polyadenylation sequence (33). In the resulting plasmid, the cDNA for Numb was flanked by the mouse Lck proximal promoter at the 5Ј end and portions of the human growth hormone gene, including introns and a polyadenylation sequence at the 3Ј end.…”

Section: Generation Of Tg Micementioning

confidence: 99%

Transgenic Expression of Numb Inhibits Notch Signaling in Immature Thymocytes But Does Not Alter T Cell Fate Specification

French

Koch

Shaye

et al. 2002

The Journal of Immunology

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The conserved adaptor protein Numb is an intrinsic cell fate determinant that functions by antagonizing Notch-mediated signal transduction. The Notch family of membrane receptors controls cell survival and cell fate determination in a variety of organ systems and species. Recent studies have identified a role for mammalian Notch-1 signals at multiple stages of T lymphocyte development. We have examined the role of mammalian Numb (mNumb) as a Notch regulator and cell fate determinant during T cell development. Transgenic overexpression of mNumb under the control of the Lck proximal promoter reduced expression of several Notch-1 target genes, indicating that mNumb antagonizes Notch-1 signaling in vivo. However, thymocyte development, cell cycle, and survival were unperturbed by mNumb overexpression, even though transgenic Numb was expressed at an early stage in thymocyte development (CD4−CD8−CD3− cells that were CD44+CD25+ or CD44−CD25+; double-negative 2/3). Moreover, bone marrow from mNumb transgenic mice showed no defects in thymopoiesis in competitive repopulation experiments. Our results suggest that mNumb functions as a Notch-1 antagonist in immature thymocytes, but that suppression of Notch-1 signaling at this stage does not alter γδ/αβ or CD4/CD8 T cell fate specification.

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“…Several tyrosine kinase mRNAs, for example lck [11] and ig/r-H [12], are known to have alternatively spliced 5'-UTR, although the function of these regions is not clear. Sequences of exons are shown in uppercase letters, introns in lower case.…”

Section: Resultsmentioning

confidence: 99%

Genomic structure and alternative splicing of the murine bhk/ctk/ntk gene

et al. 1995

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Structure of the murine lck gene and its rearrangement in a murine lymphoma cell line.

Cited by 79 publications

References 27 publications

Conserved regulation of the lymphocyte-specific expression oflckin the Fugu and mammals

Conserved regulation of the lymphocyte-specific expression oflckin the Fugu and mammals

Transgenic Expression of Numb Inhibits Notch Signaling in Immature Thymocytes But Does Not Alter T Cell Fate Specification

Genomic structure and alternative splicing of the murine bhk/ctk/ntk gene

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