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dc.contributor.authorDelgado, Elena 
dc.contributor.authorCarrera, Cristina 
dc.contributor.authorNebreda, Paloma 
dc.contributor.authorFernández-García, Aurora 
dc.contributor.authorPinilla, Milagros 
dc.contributor.authorGarcia, Valentina 
dc.contributor.authorPerez-Alvarez, Lucia 
dc.contributor.authorThomson, Michael M
dc.identifier.citationPLoS One. 2012;7(2):e30574es_ES
dc.description.abstractThe HIV-1 primary transcript undergoes a complex splicing process by which more than 40 different spliced RNAs are generated. One of the factors contributing to HIV-1 splicing complexity is the multiplicity of 3' splice sites (3'ss) used for generation of rev RNAs, with two 3'ss, A4a and A4b, being most commonly used, a third site, A4c, used less frequently, and two additional sites, A4d and A4e, reported in only two and one isolates, respectively. HIV-1 splicing has been analyzed mostly in subtype B isolates, and data on other group M clades are lacking. Here we examine splice site usage in three primary isolates of subtype C, the most prevalent clade in the HIV-1 pandemic, by using an in vitro infection assay of peripheral blood mononuclear cells. Viral spliced RNAs were identified by RT-PCR amplification using a fluorescently-labeled primer and software analyses and by cloning and sequencing the amplified products. The results revealed that splice site usage for generation of rev transcripts in subtype C differs from that reported for subtype B, with most rev RNAs using two previously unreported 3'ss, one located 7 nucleotides upstream of 3'ss A4a, designated A4f, preferentially used by two isolates, and another located 14 nucleotides upstream of 3'ss A4c, designated A4g, preferentially used by the third isolate. A new 5' splice site, designated D2a, was also identified in one virus. Usage of the newly identified splice sites is consistent with sequence features commonly found in subtype C viruses. These results show that splice site usage may differ between HIV-1 subtypes.es_ES
dc.publisherPublic Library of Sciencees_ES
dc.relation.isversionofPublisher's versiones_ES
dc.subject.meshBase Sequence es_ES
dc.subject.meshExons es_ES
dc.subject.meshGene Expression Regulation, Viral es_ES
dc.subject.meshHIV-1 es_ES
dc.subject.meshHumans es_ES
dc.subject.meshIntrons es_ES
dc.subject.meshLeukocytes, Mononuclear es_ES
dc.subject.meshMolecular Sequence Data es_ES
dc.subject.meshRNA Splice Sites es_ES
dc.subject.meshRNA Splicing es_ES
dc.subject.meshRNA, Messenger es_ES
dc.subject.meshRNA, Viral es_ES
dc.subject.meshSoftware es_ES
dc.subject.meshrev Gene Products, Human Immunodeficiency Viruses_ES
dc.titleIdentification of new splice sites used for generation of rev transcripts in human immunodeficiency virus type 1 subtype C primary isolateses_ES
dc.rights.licenseAtribución 4.0 Internacional*
dc.contributor.funderMinisterio de Ciencia e Innovación (España)es_ES
dc.identifier.journalPloS onees_ES
dc.repisalud.centroISCIII::Centro Nacional de Microbiologíaes_ES

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