The role of local secondary structure in the function of the trans-splicing motif of Brugia malayi.
A 7-nt motif (the trans-splicing motif or TSM) was previously shown to be necessary and sufficient to direct trans-splicing of transgenic mRNAs in transgenic Brugia malayi embryos. Insertion of the TSM into two genes lacking a TSM homologue resulted in trans-splicing of transgenic mRNAs from one transgene but not the other, suggesting that local sequence context might affect TSM function. To test this hypothesis, constructs inserting the TSM into different positions of two B. malayi genes were tested for their ability to support trans-splicing of transgenic mRNAs. Transgenic mRNAs derived from constructs in which the insertion of the TSM did not result in a perturbation of the local predicted secondary structure were trans-spliced, while those in which the TSM perturbed the local secondary structure were not. These data suggest that local secondary structure plays a role in the ability of the TSM to direct trans-splicing.
Global Health Infectious Disease Program, Department of Global Health, College of Public Health, University of South Florida, Tampa, FL 33612, United States.
Canhui
C
Liu
Chitra
C
Chauhan
Thomas R
TR
Unnasch
D000818:Animals; D030801:Animals, Genetically Modified; D017178:Brugia malayi / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D008958:Models, Molecular; D008967:Molecular Biology / Q000379:methods; D016254:Mutagenesis, Insertional; D009690:Nucleic Acid Conformation; D018510:RNA, Helminth / Q000737:chemistry* / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism*; D020040:Trans-Splicing
D018510:RNA, Helminth; D012333:RNA, Messenger