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Unlike EV71, virulence determinants of CA16, particularly within 5\u2032 untranslated region (5\u2032UTR), have not been investigated until now. Here, a series of nucleotides present in 5\u2032UTR of lethal but not in non-lethal CA16 strains were screened by aligning nucleotide sequences of lethal circulating Changchun CA16 and the prototype G10 as well as non-lethal SHZH05 strains. A representative infectious clone based on a lethal Changchun024 sequence and infectious mutants with various nucleotide alterations in 5\u2032UTR were constructed and further investigated by assessing virus replication <jats:italic>in vitro<\/jats:italic> and virulence in neonatal mice. Compared to the lethal infectious clone, the M2 mutant with a change from cytosine to uracil at nucleotide 104 showed weaker virulence and lower replication capacity. The predicted secondary structure of the 5\u2032UTR of CA16 RNA showed that M2 mutant located between the cloverleaf and stem-loop II, affected interactions between the 5\u2032UTR and the heterogeneous nuclear ribonucleoprotein K (hnRNP K) and A1 (hnRNP A1) that are important for translational activity. Thus, our research determined a virulence-associated site in the 5\u2032UTR of CA16, providing a crucial molecular target for antiviral drug development.<\/jats:p>","DOI":"10.1038\/srep20839","type":"journal-article","created":{"date-parts":[[2016,2,10]],"date-time":"2016-02-10T11:56:35Z","timestamp":1455105395000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Identification of a nucleotide in 5\u2032 untranslated region contributing to virus replication and virulence of Coxsackievirus A16"],"prefix":"10.1038","volume":"6","author":[{"given":"Zhaolong","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaohua","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingliang","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Hou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guanchen","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenyan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao-Fang","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,2,10]]},"reference":[{"key":"BFsrep20839_CR1","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1159\/000148922","volume":"9","author":"A Hagiwara","year":"1978","unstructured":"Hagiwara, A., Tagaya, I. & Yoneyama, T. 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