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相关概念视频

DNA Helicases00:55

DNA Helicases

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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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Condensins are large protein complexes that use ATP to fuel the assembly of chromosomes during mitosis. They transform the tangled, shapeless mass of post-interphase DNA into individualized chromosomes by compacting, organizing, and segregating chromosomal DNA.
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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
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Cohesin protein complexes are a molecular glue that holds two sister chromatids together. They play an important role both in mitosis and meiosis. In mitosis, all cohesin complexes present on the chromosomes are removed before the start of the anaphase stage.
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在Dimeric SF1 DNA螺旋中进行子单位通信.

Binh Nguyen1, John Hsieh2, Christopher J Fischer3

  • 1Department of Biochemistry & Molecular Biophysics, Washington University School of Medicine, 660 S. Euclid Ave, Saint Louis, MO 63110, USA.

Journal of molecular biology
|April 22, 2024
PubMed
概括

超级家族1基酶需要二聚化才能具有活性. 关键的DNA结合型托芬的突变取消了基酶的功能,但异构体恢复了活性,揭示了基本的子单元相互作用.

关键词:
螺旋酶激活的激活方式代表代表的代表是什么?在UvrDD中使用.功能收益的功能收益.单个分子光的光.

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科学领域:

  • 分子生物学分子生物学
  • 生物化学 生物化学
  • 酶学 是一种酶学.

背景情况:

  • 超级家族1 (SF1) 螺旋酶,如大肠杆菌Rep和UvrD,是解开DNA的必需酶.
  • 单体SF1旋酶沿单链DNA (ssDNA) 进行转位,但需要二分化才能获得完全的旋酶活性.
  • 在ssDNA结合部位中保存的托残留物对化酶功能至关重要.

研究的目的:

  • 为了研究保存的托残留物在SF1酶活性中的作用.
  • 为了阐明通过二元化激活酶激活的机制.
  • 了解Rep和UvrD异构体中的子单元通信和特异性.

主要方法:

  • 局部定向的突变生成以产生酸到氨酸突变体 (Rep(W250A,UvrD(W256A)).
  • 组合和单分子生物化学测试以测量DNA转位和酶活性.
  • 光谱法用于监测二次体内的子单元相互作用.

主要成果:

  • 在Rep和UvrD单体中,保留的托芬的突变取消了化酶活性.
  • 突变单体保留了ssDNA转位酶活性,但其速率和过程性降低了.
  • 在Rep (W250A) /wtRep和UvrD (W256A) /wtUvrD异构体中恢复了酶活性.
  • 双元体的两个子单元中的ATPase活性对于螺旋酶功能是必要的.
  • 需要子单元之间的特定相互作用,因为Rep ((W250A) 不能激活wtUvrD,反之亦然.
  • Trp光信号表明,在Rp Dimension内进行通信.

结论:

  • 保存的托芬对基酶活性至关重要,可能是通过促进基堆叠和/或适当的二聚体形成.
  • 酶的活性是通过特定的异构体的形成实现的,需要来自两个子单元的贡献.
  • 这些发现支持次单元切换机制对二维酶功能的支持,并强调了次单元间通信的重要性.