TAILIEUCHUNG - Báo cáo khoa học: Drosophila proteins involved in metabolism of uracil-DNA possess different types of nuclear localization signals

Adequate transport of large proteins that function in the nucleus is indis-pensable for cognate molecular events within this organelle. Selective pro-tein import into the nucleus requires nuclear localization signals (NLS) that are recognized by importin receptors in the cytoplasm. | ễFEBS Journal Drosophila proteins involved in metabolism of uracil-DNA possess different types of nuclear localization signals Gabor Merényi1 Emese Konya1 and Beata G. Vertessy1 2 1 Institute of Enzymology BiologicalResearch Center Hungarian Academy of Sciences Budapest Hungary 2 Department of Applied Biotechnology Budapest University of Technology and Economics Budapest Hungary Keywords cellular trafficking Drosophila melanogaster dUTPase nuclear localization signal uracil-DNA degrading factor Correspondence . Vertessy Institute of Enzymology Biological Research Center Hungarian Academy of Sciences Karolina ut 29 H-1113 Budapest Hungary Fax 36 1 466 5465 Tel 36 1 279 3116 E-mail vertessy@ Received 4 August 2009 revised 23 February 2010 accepted 1 March 2010 doi Adequate transport of large proteins that function in the nucleus is indispensable for cognate molecular events within this organelle. Selective protein import into the nucleus requires nuclear localization signals NLS that are recognized by importin receptors in the cytoplasm. Here we investigated the sequence requirements for nuclear targeting of Drosophila proteins involved in the metabolism of uracil-substituted DNA the recently identified uracil-DNA degrading factor dUTPase and the two uracil-DNA glycosylases present in Drosophila. For the uracil-DNA degrading factor NLS prediction identified two putative NLS sequences PEKRKQE 320-326 and PKRKKKR 347-353 . Truncation and site-directed mutagenesis using YFP reporter constructs showed that only one of these basic stretches is critically required for efficient nuclear localization in insect cells. This segment corresponds to the well-known prototypic NLS of SV40 T-antigen. An almost identical NLS segment is also present in the Drosophila thymine-DNA glycosylase but no NLS elements were predicted in the single-strand-specific monofunctional uracil-DNA glycosylase homolog protein. This latter protein has a .

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