Rosas-Rodríguez, Militza

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    Possible role of adenine and thymine rich sequence in the encapsidation pattern of Parvovirus LuIII
    (2007) Rosas-Rodríguez, Militza; Diffoot-Carlo, Nanette; College of Arts and Sciences - Sciences; Montalvo-Rodríguez, Rafael; Parés-Matos, Elsie I.; Department of Biology; Aponte-Huertas, María A.
    LuIII is a small, non-enveloped, icosahedral autonomous parvovirus that contains a small linear (5 kb) single stranded DNA genome and infects human cell lines lytically with no evidence of integration into cellular DNA. This parvovirus has been considered as an appropriate vector where only transient expression of a transduced gene is desired. Comparison of the LuIII sequence with those of rodent parvoviruses, minute virus of mice (MVM) and H-1 revealed that these viruses are virtually identical with respect to their genomic organization and share 80% sequence homology. However there is a 47 bp AT-rich region starting at nucleotide position 4558 that is unique to LuIII. Previous studies postulated that this AT-rich region is responsible for the symmetric encapsidation of plus and minus strand genomes by LuIII since both MVM and H-1 encapsidate primarily minus strand. To address this hypothesis, two recombinant LuIII genomes lacking the AT-rich sequence, termed pGlu883DXbaA/T- and pGlu883DXbaA/T- de novo, were transfected into HeLa cells. Southern blot analysis and probe hybridization revealed that both pGlu883DXbaA/T- constructs are unable to replicate. Cotransfection assays with LuIII minigenomes and a helper construct containing the nonstructural protein NSI under the CMV promoter (pCMVNSI) resulted in similar findings as the transfections. Thus, it is possible that the AT-rich region is a cis-acting sequence required for LuIII DNA replication.