Haaf T, Willard H F
Max Planck Institute of Molecular Genetics, Ihnestrasse 73, D-14195 Berlin, Germany.
Chromosoma. 1997 Sep;106(4):226-32. doi: 10.1007/s004120050243.
The centromeric regions of human and primate chromosomes are characterized by diverged subsets of tandemly repeated alpha-satellite DNA. Comparison of the alpha-satellites on known homologous chromosomes in human and chimpanzee provides insight into the very rapid evolution of satellite DNA sequences and the mechanisms that shape complex genomes. By using oligonucleotide primers specific for a conserved region of human alpha-satellite DNA, we have amplified a chromosome-specific alpha-satellite subset from the chimpanzee genome by the polymerase chain reaction. Fluorescence in situ hybridization showed that clones palphaPTR4N and palphaPTR4H are homologous to sequences at the centromere of the chimpanzee chromosome 4. This alpha-satellite subset is organized as a series of pentameric (higher-order) repeats, operationally defined by digestion of genomic DNA with HaeIII, MboI, RsaI, SstI, and XbaI. The lengths of four independent centromeric arrays measured by pulsed-field gel electrophoresis varied between 800 and 3,500 kb (mean = 1,850 kb, SD = 1,000 kb). Nucleotide sequence analysis demonstrated that chimpanzee chromosome 4 alpha-satellite is most closely related to the suprachromosomal subfamily II, which is evolutionarily different from the subfamily I to which the alpha-satellite on the homologous human chromosome 5 belongs. This implies that the human-chimpanzee sequence divergence has not arisen from a common ancestral alpha-satellite repeat(s) but instead represents concerted evolution of distinct repeats on homologous chromosomes.
人类和灵长类动物染色体的着丝粒区域以串联重复的α-卫星DNA的不同亚群为特征。比较人类和黑猩猩已知同源染色体上的α-卫星,有助于深入了解卫星DNA序列的快速进化以及塑造复杂基因组的机制。通过使用针对人类α-卫星DNA保守区域的寡核苷酸引物,我们利用聚合酶链反应从黑猩猩基因组中扩增出了一个染色体特异性的α-卫星亚群。荧光原位杂交显示,克隆palphaPTR4N和palphaPTR4H与黑猩猩4号染色体着丝粒处的序列同源。这个α-卫星亚群由一系列五聚体(高阶)重复序列组成,通过用HaeIII、MboI、RsaI、SstI和XbaI消化基因组DNA进行操作定义。通过脉冲场凝胶电泳测量的四个独立着丝粒阵列的长度在800至3500 kb之间变化(平均值 = 1850 kb,标准差 = 1000 kb)。核苷酸序列分析表明,黑猩猩4号染色体α-卫星与超染色体亚家族II关系最为密切,后者在进化上不同于同源人类5号染色体上的α-卫星所属的亚家族I。这意味着人类与黑猩猩的序列差异并非源于共同的祖先α-卫星重复序列,而是代表了同源染色体上不同重复序列的协同进化。