Pearson C E, Eichler E E, Lorenzetti D, Kramer S F, Zoghbi H Y, Nelson D L, Sinden R R
Center for Genome Research, Institute of Biosciences and Technology, Department of Biochemistry and Biophysics, Texas A&M University, Houston, Texas, 77030, USA.
Biochemistry. 1998 Feb 24;37(8):2701-8. doi: 10.1021/bi972546c.
Models for the disease-associated expansion of trinucleotide repeats involve the participation of alternative DNA structures during replication, repair, or recombination. CAT or AGG interruptions within the (CAG)n or (CGG)n repeats of SCA1 or FRAXA, respectively, confer increased genetic stability to the repeats. In this study, we report the formation of slipped strand structures (S-DNA) using genomic sequences containing pure and interrupted SCA1 and FRAXA repeats having lengths above and below the genetic stability thresholds. S-DNA forms within the repeats during annealing of complementary strands containing equal lengths of repeats. Increased lengths of pure repeats led to an increased propensity for S-DNA formation. CAT or AGG interruptions have both quantitative and qualitative effects upon S-DNA formation: they decrease the total amount of slipped structures as well as limit the specific isomers formed. This demonstrates a unifying inhibitory effect of interruptions in both (CAG)n and (CGG)n tracts. We also present transmission stability data for SCA1 and FRAXA alleles spanning the thresholds and compare these with the ability to form slipped structures. The effect of both the length and purity of the repeat tract on the propensity of slipped structure formation correlates with their effect on genetic instability and disease, suggesting that S-DNA structures may be models for mutagenic intermediates in instability.
三核苷酸重复序列疾病相关扩增的模型涉及复制、修复或重组过程中替代DNA结构的参与。分别位于脊髓小脑共济失调1型(SCA1)的(CAG)n重复序列或脆性X综合征A(FRAXA)的(CGG)n重复序列中的CAT或AGG中断,赋予这些重复序列更高的遗传稳定性。在本研究中,我们报告了使用包含纯合和中断的SCA1及FRAXA重复序列的基因组序列形成滑链结构(S-DNA),这些重复序列的长度高于和低于遗传稳定性阈值。在含有等长重复序列的互补链退火过程中,重复序列内会形成S-DNA。纯合重复序列长度增加导致S-DNA形成倾向增加。CAT或AGG中断对S-DNA形成具有定量和定性影响:它们减少了滑链结构的总量,并限制了形成的特定异构体。这证明了(CAG)n和(CGG)n序列中断具有统一的抑制作用。我们还展示了跨越阈值的SCA1和FRAXA等位基因的传递稳定性数据,并将其与形成滑链结构的能力进行比较。重复序列的长度和纯度对滑链结构形成倾向的影响与其对遗传不稳定性和疾病的影响相关,这表明S-DNA结构可能是不稳定性中诱变中间体的模型。