Boán F, Rodríguez J M, Gómez-Márquez J
Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Santiago, 15706 Santiago de Compostela, La Coruña, Galicia, Spain.
J Mol Biol. 1998 May 8;278(3):499-505. doi: 10.1006/jmbi.1998.1714.
Several features indicate that the low polymorphic human minisatellite MsH42 region could be involved in recombination. It contains different well-known recombination motifs, is able to generate single-stranded loops and is specifically recognized by nuclear proteins. These characteristics led us to investigate the possible recombinogenic activity of the MsH42 region in terms of intramolecular recombination. We constructed two plasmids, one of them carrying two copies of the minisatellite region and the other one containing sequences upstream of this repetitive region. We showed that MsH42 strongly stimulates intramolecular in vitro recombination, approximately 22 times more than the control sequence, solely when the source of biological extract is mouse testes, suggesting that MsH42 could be a hotspot involved in meiotic recombination. Furthermore, there is a direct relationship between the frequency of equal crossovers and the enhancement of recombination. Interestingly, the third repeat of the minisatellite array is always involved in the resolution of unequal crossovers leading to minisatellite shortening. As far as we know, our results provide the first evidence that a non-hypervariable minisatellite can enhance homologous recombination.
若干特征表明,低多态性的人类小卫星MsH42区域可能参与重组。它包含不同的知名重组基序,能够产生单链环,并且能被核蛋白特异性识别。这些特征促使我们从分子内重组的角度研究MsH42区域可能的重组活性。我们构建了两个质粒,其中一个携带小卫星区域的两个拷贝,另一个包含该重复区域上游的序列。我们发现,仅当生物提取物的来源是小鼠睾丸时,MsH42能强烈刺激分子内体外重组,比对照序列大约多22倍,这表明MsH42可能是参与减数分裂重组的热点区域。此外,均等交叉的频率与重组增强之间存在直接关系。有趣的是,小卫星阵列的第三个重复序列总是参与导致小卫星缩短的不等交叉的解析。据我们所知,我们的结果首次证明了一个非高变小卫星可以增强同源重组。