Biochemical characterization of two Cdc6/ORC1-like proteins from the crenarchaeon Sulfolobus solfataricus
Biochemical characterization of two Cdc6/ORC1-like proteins from the crenarchaeon Sulfolobus solfataricus
复制标题
DOI：
10.1007/s00792-005-0473-0
复制
发表时间：
2006-02-01
期刊：
影响因子：
2.9
通讯作者：
Pisani, FM
中科院分区：
文献类型：
--
作者：
De Felice, M;Esposito, L;Pisani, FM
古细菌蛋白质的生物学作用，同源的真核复制起始因子细胞分裂控制（Cdc 6）和起源识别复合物（ORC 1），尚未明确建立。极端嗜热泉古菌Sulfolobus solfataricus（简称Sso）具有3个Cdc 6/ORC 1样因子，分别命名为Sso Cdc 6 -1、Cdc 6 -2和Cdc 6 -3。本研究是对Sso Cdc 6 -1和Cdc 6 -3的生化特性的报告。通过凝胶过滤分析发现，Sso Cdc 6 -1或Cdc 6 -3在溶液中表现为单体。这两种因子都能与各种单链和双链DNA配体结合，但Sso Cdc 6 -3显示出更高的DNA结合亲和力。还观察到Sso Cdc 6 -1或Cdc 6 -3抑制S. solfataricus homo-hexameric mini-chromosome maintenance（MCM）-like DNA解旋酶（Sso MCM）;尽管它们强烈刺激Sso MCM与含气泡的合成寡核苷酸的相互作用。该研究还表明，通过表面等离子体共振测量，Sso Cdc 6 -2与Sso Cdc 6 -1或Sso Cdc 6 -3发生物理相互作用。这些发现可能提供重要的线索，需要了解这三个Cdc 6因子中的每一个在S.苏氏细胞
The biological role of archaeal proteins, homologous to the eukaryal replication initiation factors of cell division control (Cdc6) and origin recognition complex (ORC1), has not yet been clearly established. The hyperthermophilic crenarchaeon Sulfolobus solfataricus (referred to as Sso) possesses three Cdc6/ORC1-like factors, which are named Sso Cdc6-1, Cdc6-2 and Cdc6-3. This study is a report on the biochemical characterization of Sso Cdc6-1 and Cdc6-3. It has been found that either Sso Cdc6-1 or Cdc6-3 behave as monomers in solutions by gel filtration analyses. Both factors are able to bind to various single-stranded and double-stranded DNA ligands, but Sso Cdc6-3 shows a higher DNA-binding affinity. It has also been observed that either Sso Cdc6-1 or Cdc6-3 inhibit the DNA unwinding activity of the S. solfataricus homo-hexameric mini-chromosome maintenance (MCM)-like DNA helicase (Sso MCM); although they Strongly stimulate the interaction of the Sso MCM with bubblecontaining synthetic oligonucleotides. The Study has also showed, With Surface plasmon resonance measurements, that Sso Cdc6-2 physically interacts with either Sso Cdc6-1 or Sso Cdc6-3. These findings may provide important Clues needed to understand the biological role that is played by each of these three Cdc6 factors during the DNA replication initiation process in the S. solfataricus cells.