Leeck C L, Forney J D
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907-1153.
J Biol Chem. 1994 Dec 9;269(49):31283-8.
Paramecium tetraurelia stock 51 can express at least 11 different surface antigens, yet only one type is found on the surface of a cell at any given time. The mechanism that controls this mutually exclusive expression is unknown. A previous study has shown that the 51A surface antigen gene is regulated at the level of transcription (Gilley, D., Rudman, B. M., Preer, J. R., Jr., and Polisky, B. (1990) Mol. Cell. Biol. 10, 1538-1544). We show here that the 51B surface antigen gene is also transcriptionally regulated, and when the 51A and 51B genes are cotransformed into an A-, B- mutant, the 51A antigen is dominant at 27 degrees C just as in wild type cells. We have utilized this cotransformation system to experimentally determine that 273 base pairs of DNA upstream of the 51A gene is sufficient to allow the dominant expression of A, but 150 base pairs is not adequate. A hybrid gene that contains the upstream region of 51B attached to the 51A transcribed region was cotransformed with the 51B gene into the A-, B- mutant. Despite containing the same upstream sequences, only the hybrid 51B/A was transcribed at 27 degrees C. These results suggest the upstream region is required but not sufficient for mutually exclusive transcriptional control.
四膜虫51号品系能够表达至少11种不同的表面抗原,但在任何给定时间,细胞表面仅发现一种类型的抗原。控制这种互斥表达的机制尚不清楚。先前的一项研究表明,51A表面抗原基因在转录水平受到调控(吉利,D.,鲁德曼,B.M.,普里尔,J.R. Jr.,和波利斯基,B.(1990年)《分子与细胞生物学》10,1538 - 1544)。我们在此表明,51B表面抗原基因同样受到转录调控,并且当51A和51B基因共转化到A - 、B - 突变体中时,51A抗原在27摄氏度时如同在野生型细胞中一样占主导地位。我们利用这个共转化系统通过实验确定,51A基因上游273个碱基对的DNA足以使A抗原占主导表达,但150个碱基对则不够。一个包含与51A转录区域相连的51B上游区域的杂合基因与51B基因一起共转化到A - 、B - 突变体中。尽管包含相同的上游序列,但只有杂合的51B/A在2q7摄氏度时被转录。这些结果表明,上游区域对于互斥转录控制是必需的,但并不充分。