Bressan Debra A, Vazquez Julio, Haber James E
Rosenstiel Center and Department of Biology, Brandeis University, Waltham, MA 02454-9910, USA.
J Cell Biol. 2004 Feb 2;164(3):361-71. doi: 10.1083/jcb.200311063. Epub 2004 Jan 26.
Mating-type gene (MAT) switching in budding yeast exhibits donor preference. MATa preferentially recombines with HML near the left telomere of chromosome III, whereas MATalpha prefers HMR near the right telomere. Donor preference is controlled by the recombination enhancer (RE) located proximal to HML. To test if HML is constrained in pairing with MATalpha, we examined live-cell mobility of LacI-GFP-bound lactose operator (lacO) arrays inserted at different chromosomal sites. Without induction of recombination, lacO sequences adjacent to HML are strongly constrained in both MATalpha and RE-deleted MATa strains, compared with MATa. In contrast, chromosome movement at HMR or near a telomere of chromosome V is mating-type independent. HML is more constrained in MATa Deltare and less constrained in MATa RE+ compared with other sites. Although HML and MATa are not prealigned before inducing recombination, the three-dimensional configuration of MAT, HML, and HMR is mating-type dependent. These data suggest there is constitutive tethering of HML, which is relieved in MATa cells through the action of RE.
芽殖酵母中的交配型基因(MAT)转换表现出供体偏好。MATa优先与位于第三条染色体左端粒附近的HML重组,而MATα则偏好位于右端粒附近的HMR。供体偏好由位于HML近端的重组增强子(RE)控制。为了测试HML与MATα配对是否受到限制,我们检测了插入不同染色体位点的与LacI-GFP结合的乳糖操纵子(lacO)阵列在活细胞中的移动性。在未诱导重组的情况下,与MATa相比,在MATα和RE缺失的MATa菌株中,与HML相邻的lacO序列都受到强烈限制。相比之下,HMR或第五条染色体端粒附近的染色体移动与交配型无关。与其他位点相比,HML在MATa Deltare中受到的限制更大,而在MATa RE+中受到的限制较小。尽管在诱导重组之前HML和MATa没有预先对齐,但MAT、HML和HMR的三维构型是交配型依赖的。这些数据表明存在HML的组成型束缚,在MATa细胞中通过RE的作用这种束缚得以解除。