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    The T4 TerL prohead packaging motor does not drive DNA translocation by a proposed dehydration mechanism

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    Author
    Black, L.W.
    Yan, B.
    Ray, K.
    Date
    2020
    Journal
    Viruses
    Publisher
    MDPI AG
    Type
    Article
    
    Metadata
    Show full item record
    See at
    https://doi.org/10.3390/v12050522
    Abstract
    A "DNA crunching" linear motor mechanism that employs a grip-and-release transient spring like compression of B- to A-form DNA has been found in our previous studies. Our FRET measurements in vitro show a decrease in distance from TerL to portal during packaging; furthermore, there is a decrease in distance between closely positioned dye pairs in the Y-stem of translocating Y-DNA that conforms to B- and A- structure. In normal translocation into the prohead the TerL motor expels all B-form tightly binding YOYO-1 dye that cannot bind A-form. The TerL motor cannot package A-form dsRNA. Our work reported here shows that addition of helper B form DNA:DNA (D:D) 20mers allows increased packaging of heteroduplex A-form DNA:RNA 20mers (D:R), evidence for a B- to A-form spring motor pushing duplex nucleic acid. A-form DNA:RNA 25mers, 30mers, and 35mers alone are efficiently packaged into proheads by the TerL motor showing that a proposed hypothetical dehydration motor mechanism operating on duplex substrates does not provide the packaging motor force. Taken together with our previous studies showing TerL motor protein motion toward the portal during DNA packaging, our present studies of short D:D and D:R duplex nucleic acid substrates strongly supports our previous evidence that the protein motor pushes rather than pulls or dehydrates duplex substrates to provide the translocation into prohead packaging force. Copyright 2020 by the authors.
    Sponsors
    Funding: Research reported in this publication was supported by the National Institute of General Medical Sciences and National Institute of Allergy and Infectious Diseases at the National Institutes of Health under Award Numbers R01 GM118766 and R01 AI150447.
    Keyword
    DNA crunching
    Heteroduplex A-form DNA:RNA packaging
    Linear motor mechanism
    T4 TerL prohead
    Identifier to cite or link to this item
    https://www.scopus.com/inward/record.uri?eid=2-s2.0-85084579795&doi=10.3390%2fv12050522&partnerID=40&md5=0c17e0491d25904f3cd842382d78beb1; http://hdl.handle.net/10713/12832
    ae974a485f413a2113503eed53cd6c53
    10.3390/v12050522
    Scopus Count
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    UMB Open Access Articles 2020

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