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Analysis of Paramecium tetraurelia A-51 Surface Antigen Gene Mutants Reveals Positive-Feedback Mechanisms for Maintenance of Expression and Temperature-Induced Activation

Department of Biochemistry, Purdue University, 175 S. University Street, West Lafayette, Indiana 47907-2063

Received 18 April 2005/ Accepted 8 August 2005

In Paramecium tetraurelia, variable surface antigen loci show mutually exclusive expression which is controlled primarily at the transcriptional level. Clonally stable expression of a single antigen has attracted models involving self-regulation by their gene products. However, direct demonstration of self-feedback at the molecular level has been complicated due to the inability to separate the functional gene from its product as well as copy number effects associated with injected extrachromosomal DNA in the polygenomic somatic nucleus. In this study, we exploited several germ line termination and frameshift mutations in the A-51 surface antigen gene to analyze variable surface antigen expression. These mutant alleles have the same copy number as the wild-type allele and therefore eliminate possible copy number effects. The mutant alleles were not transcribed at 27°C, consistent with positive-feedback models for gene expression. However, further analysis showed that high temperatures (34°C) induced transcription of the mutant A genes even in the presence of a different antigen on the cell surface. Thus, transcription was temperature dependent. Unlike wild-type cells, transcription of the mutant A genes at high temperatures was not maintained after temperature shift back to 27°C in homozygous mutant cells. Importantly, transcription of the mutant allele was maintained at 27°C in heterozygous cells with one copy of the wild-type allele. These results indicate that expression of the wild-type gene is required to stabilize its own transcriptional state at 27°C.