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DNA helicases were discovered in E. The common function of helicases accounts for the fact that they display a certain degree of amino acid sequence homology ; they all possess sequence motifs located in the interior of their primary structure , involved in ATP binding, ATP hydrolysis and translocation along the nucleic acid substrate.

The variable portion of the amino acid sequence is related to the specific features of each helicase. The presence of these helicase motifs allows putative helicase activity to be attributed to a given protein, but does not necessarily confirm it as an active helicase.

Conserved motifs do, however, support an evolutionary homology among enzymes. Based on these helicase motifs, a number of helicase superfamilies have been distinguished.

Helicases are classified in 6 groups superfamilies based on their shared sequence motifs. They are also classified by translocation polarity.

Various types of mutations found in ATRX have been found to be associated with ATR-X, including most commonly single-base missense mutations, as well as nonsense, frameshift, and deletion mutations.

XPD helicase mutations leading to trichothiodystrophy are found throughout the protein in various locations involved in protein-protein interactions.

It has been suggested that XPD helicase mutations leading to Cockayne syndrome could be the result of mutations within XPD, causing rigidity of the protein and subsequent inability to switch from repair functions to transcription functions due to a "locking" in repair mode.

Bloom syndrome is characterized by a predisposition to cancer with early onset, with a mean age-of-onset of 24 years.

Werner syndrome is a disorder of premature aging, with symptoms including early onset of atherosclerosis and osteoporosis and other age related diseases, a high occurrence of sarcoma, and death often occurring from myocardial infarction or cancer in the 4th to 6th decade of life.

Rothmund-Thomson syndrome, also known as poikiloderma congenitale , is characterized by premature aging, skin and skeletal abnormalities, rash, poikiloderma , juvenile cataracts, and a predisposition to cancers such as osteosarcomas.

During meiosis DNA double-strand breaks and other DNA damages in a chromatid are repaired by homologous recombination using either the sister chromatid or a homologous non-sister chromatid as template.

It was suggested that COs are restricted because of the long term costs of CO recombination, that is, the breaking up of favorable genetic combinations of alleles built up by past natural selection.

They also play an important role in sensing viral RNAs. On the other hand, ring-forming RNA helicases have been found in bacteria and viruses.

Although these proteins carry the typical helicase motifs, hydrolize ATP in a nucleic acid-dependent manner, and are built around a helicase core, in general, no unwinding activity is observed.

RNA helicases that do exhibit unwinding activity have been characterized by at least two different mechanisms: Canonical duplex unwinding is the stepwise directional separation of a duplex strand, as described above, for DNA unwinding.

However, local strand separation occurs by a process wherein the helicase enzyme is loaded at any place along the duplex.

This is usually aided by a single-strand region of the RNA, and the loading of the enzyme is accompanied with ATP binding. An RNA helicase database [62] is currently available online that contains a comprehensive list of RNA helicases with information such as sequence, structure, and biochemical and cellular functions.

Various methods are used to measure helicase activity in vitro. These methods range from assays that are qualitative assays that usually entail results that do not involve values or measurements to quantitative assays with numerical results that can be utilized in statistical and numerical analysis.

In , the first direct biochemical assay was developed for measuring helicase activity. Other methods were later developed that incorporated some, if not all of the following: Commercially available diagnostic kits are also available.

One such kit is the "Trupoint" diagnostic assay from PerkinElmer , Inc. This assay is a time-resolved fluorescence quenching assay that utilizes the PerkinEmer "SignalClimb" technology that is based on two labels that bind in close proximity to one another but on opposite DNA strands.

The other label is an organic quencher molecule. The basis of this assay is the "quenching" or repressing of the lanthanide chelate signal by the organic quencher molecule when the two are in close proximity - as they would be when the DNA duplex is in its native state.

Upon helicase activity on the duplex, the quencher and lanthanide labels get separated as the DNA is unwound. This loss in proximity negates the quenchers ability to repress the lanthanide signal, causing a detectable increase in fluorescence that is representative of the amount of unwound DNA and can be used as a quantifiable measurement of helicase activity.

The execution and use of single-molecule fluorescence imaging techniques, focusing on methods that include optical trapping in conjunction with epifluorescent imaging, and also surface immobilization in conjunction with total internal reflection fluorescence visualization.

Combined with microchannel flow cells and microfluidic control, allow individual fluorescently labeled protein and DNA molecules to be imaged and tracked, affording measurement of DNA unwinding and translocation at single-molecule resolution.

This determination of polarity is vital in f. From Wikipedia, the free encyclopedia. Journal of Biological Chemistry.

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