The 5′ Coding Region of Paramecium Surface Antigen Genes Controls Mutually Exclusive Transcription

• Published in: Proceedings of the National Academy of Sciences - PNAS 1996-04, Vol.93 (7), p.2838-2843
• Main Authors: Leeck, Charles L., Forney, James D.
• Format: Article
• Language: English
• Subjects: Animals; Antigens, Protozoan - biosynthesis; Antigens, Protozoan - genetics; Antigens, Surface - biosynthesis; Antigens, Surface - genetics; Base Sequence; Biochemistry; Cell lines; Cell Nucleus - metabolism; DNA; DNA Primers; DNA Probes; Gene Expression Regulation; Genes; Genes, Protozoan; Genetics; Molecular Sequence Data; Nucleotides; Open reading frames; Paramecium tetraurelia; Paramecium tetraurelia - genetics; Paramecium tetraurelia - metabolism; Plasmids; Polymerase Chain Reaction; Protein Biosynthesis; Regulatory Sequences, Nucleic Acid; RNA; RNA, Protozoan - biosynthesis; Surface antigens; Transcription, Genetic; Transformed cell line; Untranslated regions
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.7.2838

Paramecium tetraurelia stock 51 can express at least 11 different types of surface antigens, yet only a single type is expressed on the surface of an individual cell at any one time. The differential expression of stock 51 type A and B surface antigen genes (51A and 51B) is regulated at the level of transcription. Previously, we reported that nucleotide sequences upstream of position -26 (relative to the start of translation) in the 51A and 51B surface antigen genes are necessary for transcriptional activity but are not sufficient to direct differential transcriptional control. In this report we demonstrate that at least some of the critical elements necessary for differential transcription of the 51A and 51B genes lie within the 5′ coding region. A hybrid gene that contains 51B upstream sequences (-475 to +1) attached to the ATG start codon of 51A is not cotranscribed with the 51B gene. In contrast, further substitution with 51B sequences (-1647 to +885) allows the chimeric gene to be coexpressed with 51B. A different hybrid gene containing a substitution of 51B sequence from -26 to +885 in the 51A gene is also coexpressed with 51B, revealing that the critical elements within the coding region of 51B do not require 51B upstream sequences for their effect. Coinjection of the 51A gene with the chimeric gene that contains 51B up to +885 showed that the same sequences that allow coexpression with 51B prevent cotranscription with 51A. Together, these results demonstrate that a region downstream of the transcriptional start site between nucleotide positions +1 and +885 (relative to translational start) is necessary to control differential transcriptional activity.