NOA36 Protein Contains a Highly Conserved Nucleolar Localization Signal Capable of Directing Functional Proteins to the Nucleolus, in Mammalian Cells

de Melo, Ivan S.; Jimenez-Nuñez, Maria D.; Iglesias, Concepción; Campos-Caro, Antonio; Moreno-Sanchez, David; Ruiz, Felix A.; Bolívar, Jorge

KKLKKRNK [16] P14ARF RRGAQLRRPRHSHPTRARRCP [17] SVV-EX3 MQRKPTIRRKNLRKLRRK [15] NIK RKKRKKK [18] hLa QESLNKWKSKGRRFKGKGKGNKAAQPGSGKGK [19] ING-1 MTPKEKKAKTSKKKKRSKAKA [20] NOLP RKRIRTYLKSCRRMKRSGFEMSRPIPSHLT [21] IGF-I GTEASLQIRGKKKEQRREIGSRNAECRGKKGK [22] Werner VSRYNKFMKICALTKKGRNWLHKANTES [23] Nucleolin RGGGGGGGDFKPQGKKTKFR [24] Parafibromin RRAATENIPVVRRPDRK-(X) 301 -KKQGCQRENETLIQRRK [25] PTHrP GKKKKGKPGKRREQEKKKRRT [26] DEDD KRPARGRATLGSQPKRRKSV [27] Angiogenin IMRRRGL [28] Protein Alpha RRRANNRRR [29] MEQ RRRKRNRDAARRRRRKQ [30] Tat GRKKRRQRRAP [31] Rev RQARRNRRRRWERQR [32] Rex PKTRRRPRRSQRKRP [33] I14L MSRRNKRSRRRRKKPLNTIQ [34] These clusters of basic amino acids can be found in human proteins as MDM2 [16] , p14ARF [17] , SVV-EX3 [15] , NIK [18] , hLa [19] , ING-I [20] , NOLP [21] , IGF-I [22] , Werner [23] , Nucleolin [24] , Parafibromin [25] , PTHrP [26] , DEDD [27] , Angiogenin [28] and in several viral proteins such as Protein alpha [29] , MEQ (protein of Marek's disease virus (MDV) [30] , Tat [31] and Rev [32] (both proteins of human immunodeficiency virus type 1), Rex (protein of human T lymphotropic virus type [33] , I14L (protein of African swine fever virus) [34] . (Clontech, Mountainview, CA). As a control, the coding sequence of scPPX1 was fused directly with the Flag tag and cloned into the eukaryotic expression vector pIRES2-EGFP. Constructs were verified by sequencing (Servicio Central de Ciencia y Tecnología, University of Cadiz). Sequences of oligonucleotides and mutagenesis primers are available on request.