Disruption of the Expression of a Non-Coding RNA Significantly Impairs Cellular Differentiation in Toxoplasma gondii.docVIP

Int. J. Mol. Sci. 2013, 14, 611-624; doi:10.3390/ijmOPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms Article Disruption of the Expression of a Non-Coding RNA Significantly Impairs Cellular Differentiation in Toxoplasma gondii Veerupaxagouda Patil, Pamela J. Lescault, Dario Lirussi, Ann B. Thompson and Mariana Matrajt * Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Dr. Burlington, VT 05405, USA; E-Mails: vkpatil13@ (V.P.); pamela.lescault@ (P.J.L.); dlirussi@ (D.L.); abthomps@ (A.B.T.) * Author to whom correspondence should be addressed; E-Mail: mmatrajt@; Tel.: +1-802-656-3671; Fax: +1-802-656-8749. Received: 2 November 2012; in revised form: 14 December 2012 / Accepted: 18 December 2012 / Published: 28 December 2012 Abstract: The protozoan parasite Toxoplasma gondii is an important human and veterinary pathogen. Asexual replication of T. gondii in humans and intermediate hosts is characterized by two forms: rapidly growing “tachyzoites” and latent “bradyzoite” tissue cysts. Tachyzoites are responsible for acute illness and congenital neurological birth defects, while the more slowly dividing bradyzoite form can remain latent within the tissues for many years, representing a threat to immunocompromised patients. We have developed a genetic screen to identify regulatory genes that control parasite differentiation and have isolated mutants that fail to convert to bradyzoites. One of these mutants has an insertion disrupting a locus that encodes a developmentally regulated non-coding RNA transcript, named Tg-ncRNA-1. Microarray hybridizations suggest that Tg-ncRNA-1 is involved in the early steps of bradyzoite differentiation. Since Tg-ncRNA-1 does not contain an open reading frame, we used the algorithm Coding Potential Calculator (CPC) that evaluates the protein-coding potential of