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Accurate 5 Splice-Site Selection in Mouse κ Immunoglobulin Light Chain Premessenger RNAs is not Cell-Type-Specific

Dean H. Kedes and Joan A. Steitz
Proceedings of the National Academy of Sciences of the United States of America
Vol. 84, No. 22 (Nov. 15, 1987), pp. 7928-7932
Stable URL: http://www.jstor.org/stable/30574
Page Count: 5
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Accurate 5′ Splice-Site Selection in Mouse κ  Immunoglobulin Light Chain Premessenger RNAs is not Cell-Type-Specific
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Abstract

In mature mouse B lymphocytes, immunoglobulin κ light chain transcripts contain an intervening sequence separating the recombined variable (V) plus joining (J) exon from the distant constant (C) exon. After V--J recombination, this intervening sequence can include as many as three unused but very similar J-region 5 splice sites. Each of these sites is potentially functional if the gene is appropriately recombined. It is unclear how the splicing machinery distinguishes among these 5 splice sites, always choosing the most upstream site. We used synthetic transcripts of κ gene sequences containing J3 and J4 in both the germ-line and the recombined configurations to study the pattern of 5 splice-site selection in vitro. We find that both HeLa cell and lymphocyte nuclear extracts fail to discriminate between the J3- and J4-region 5 splice sites. In contrast, after transfection into HeLa cells, similar κ light chain transcripts are spliced correctly at the most upstream 5 splice site--that which is used in κ -producing cells. We conclude that accurate 5 splice-site selection in the mouse κ light chain is neither cell-type- nor species-specific. Potential mechanisms for this controlling step in gene expression are discussed.

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