Multiple Features Contribute to the Use of the Immunoglobulin M Secretion-Specific Poly(A) Signal but Are Not Required for Developmental Regulation

ABSTRACT The secretory-specific poly(A) signal (μs) of the immunoglobulin μ gene plays a central role in regulating alternative RNA processing to produce RNAs that encode membrane-associated and secreted immunoglobulins. This poly(A) signal is in direct competition with a splice reaction, and regulation requires that these two reaction efficiencies be balanced. The μs poly(A) signal has several unique sequence features that may contribute to its strength and regulation. Site-directed mutations and small internal deletions made in the intact μ gene show that an extensive AU/A-rich sequence surrounding AAUAAA enhances signal use and that, of the two potential downstream GU-rich elements, both of which appear suboptimally located, only the proximal GU-rich sequence contributes substantially to use of this signal. A GU-rich sequence placed at a more standard location did not improve μs poly(A) signal use. All μ genes tested that contained modified μs poly(A) signals were developmentally regulated, indicating that the GU-rich sequences, the sequences between them previously identified as suboptimal U1A binding sites, and an upstream suboptimal U1A site do not contribute to μ mRNA processing regulation. Expression of wild-type and modified μ genes in HeLa cells overexpressing U1A also failed to demonstrate that U1A contributes to μs poly(A) signal regulation.