Abstract

The human CD45 gene encodes five isoforms of a transmembrane tyrosine phosphatase that differ in their extracellular domains as a result of alternative splicing of exons 4-6. Expression of the CD45 isoforms is tightly regulated in peripheral T cells such that resting cells predominantly express the larger CD45 isoforms, encoded by mRNAs containing two or three variable exons. In contrast, activated T cells express CD45 isoforms encoded by mRNAs lacking most or all of the variable exons. We have previously identified the sequences within CD45 variable exon 4 that control its level of inclusion into spliced mRNAs. Here we map the splicingregulatory sequences within CD45 variable exons 5 and 6. We show that, like exon 4, exons 5 and 6 each contain an exonic splicing silencer (ESS) and an exonic splicing enhancer (ESE), which together determine the level of exon inclusion in naïve cells. We further demonstrate that the primary activation-responsive silencing motif in exons 5 and 6 is homologous to that in exon 4 and, as in exon 4, binds specifically to the protein heterogeneous nuclear ribonucleoprotein L. Together these studies reveal common themes in the regulation of the CD45 variable exons and provide a mechanistic explanation for the observed physiological expression of CD45 isoforms.

Highlights

  • The importance of regulated alternative splicing in humans is exemplified by the CD45 gene, which encodes a transmembrane proteintyrosine phosphatase [3]

  • Variable exons 4 and 6 are only included in a fraction of mRNAs expressed in resting T cells, whereas exon 5 is expressed in almost all of these mRNAs [10]

  • Whereas the inclusion of exons 4 and 6 is clearly repressed upon T cell activation, it has been debated whether the splicing of exon 5 is inherently regulated or is a stochastic event influenced by the neighboring effects of the other two variable exons [3]

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Summary

Introduction

The importance of regulated alternative splicing in humans is exemplified by the CD45 gene, which encodes a transmembrane proteintyrosine phosphatase [3]. We have identified the regulatory sequences that control inclusion of CD45 variable exon 4, including a motif referred to as the activation-responsive sequence (ARS) consensus that is required for exon silencing upon T cell activation [18, 19].

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