Hollingsworth R E, Sclafani R A
Department of Biochemistry, Biophysics, and Genetics, University of Colorado Health Sciences Center, Denver 80262.
Chromosoma. 1993 Jun;102(6):415-20. doi: 10.1007/BF00360406.
In budding yeast, mitotic DNA replication initiates at sequence-specific replication origins, the prototype for which is ARS1. Initiation serves as the primary control point for mitotic DNA replication, and is catalyzed by the Cdc7 protein kinase. In contrast, premeiotic DNA replication apparently does not require Cdc7, and the existence and nature of specific replication origins in the meiotic division cycle have not been previously reported. We have begun to investigate the mechanism of premeiotic DNA synthesis by determining whether or not ARS1 functions as a DNA replication origin in meiosis. We have taken advantage of the fact that transcription through ARS1 disrupts its ability to function as an origin to show that ARS1 is required for premeiotic DNA replication of a plasmid bearing this element. Further, premeiotic replication from ARS1 still occurs in a cdc7 mutant strain held at conditions non-permissive for Cdc7 protein kinase activity. These findings reveal that premeiotic DNA replication can initiate from origins also used in mitosis, and is not regulated by Cdc7. Taken together with previous findings implicating Cdc7 in meiotic DNA recombination and induced mutagenesis, these findings prompt us to postulate that the Cdc7 protein kinase regulates some step common to several DNA metabolic processes such as local disassembly of chromatin or activation of a key component of the DNA metabolic machinery.
在出芽酵母中,有丝分裂DNA复制起始于序列特异性复制起点,其原型是ARS1。起始过程是有丝分裂DNA复制的主要控制点,由Cdc7蛋白激酶催化。相比之下,减数分裂前的DNA复制显然不需要Cdc7,并且此前尚未报道减数分裂周期中特异性复制起点的存在及性质。我们已开始通过确定ARS1在减数分裂中是否作为DNA复制起点来研究减数分裂前DNA合成的机制。我们利用了这样一个事实,即通过ARS1的转录会破坏其作为起点的功能,以此表明ARS1对于携带该元件的质粒的减数分裂前DNA复制是必需的。此外,在处于不允许Cdc7蛋白激酶活性的条件下的cdc7突变菌株中,ARS1的减数分裂前复制仍然会发生。这些发现揭示了减数分裂前DNA复制可以从有丝分裂中也使用的起点起始,并且不受Cdc7的调控。结合先前将Cdc7与减数分裂DNA重组和诱导诱变相关联的发现,这些发现促使我们推测Cdc7蛋白激酶调节几个DNA代谢过程共有的某个步骤,例如染色质的局部解聚或DNA代谢机制关键组分的激活。