Biopolym. Cell. 2010; 26(2):115-120.
Structure and Function of Biopolymers
Identification and functional analysis of an alternative promoter of human intersectin 1 gene
1Kropyvko S. V., 1Tsyba L. O., 1Skrypkina I. Ya., 1Rynditch A. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


Intersectin 1 (ITSN1) gene encodes an evolutionarily conserved adaptor protein that functions in clathrin-mediated endocytosis, cell signalling, apoptosis and cytoskeleton rearrangements. Its expression is characterized by multiple alternative splicing. Alternative promoter usage is an additional way to create diversity and flexibility in the regulation of gene expression. The aim of this study was to identify possible alternative promoters of ITSN1 gene. Methods. In silico prediction, 5' RACE, RT-PCR and reporter gene expression assay were used for identification and functional characterization of alternative promoter region. Results. We detected an alternative promoter of human ITSN1 gene which is located in intron 5 and generates 5' truncated transcripts containing in-frame ATG codon with strong Kozak sequence and could encode an N-terminally truncated isoforms lacking first EH domain. The region located 246–190 bp upstream of exon 6 is required for alternative promoter activity. ITSN1 transcripts generated from an alternative promoter were detected in human kidney, liver, lung and brain tissues. However, the level of their expression was significantly lower than that of major ITSN1 isoforms. Conclusion. The results obtained suggest that alternative promoter region located in intron 5 of ITSN1 gene functions as a weak promoter. Further experiments are required to clarify the role of 5' truncated ITSN1 transcripts.
Keywords: intersectin 1, alternative promoter, alternative splicing, 5' UTR, adaptor proteins


[1] Pechstein A., Shupliakov O., Haucke V. Intersectin 1: a versatile actor in the synaptic vesicle cycle Biochem. Soc. Trans 010 38, Pt 1 P. 181–186.
[2] Tsyba L. O., Skrypkina I. Ya., Nikolaienko O. V., Ferenent G. O., Gordiyuk V. V., Gardiner K., Rynditch A. V. Alternative splicing of intersectin 1 gene pre-mRNA: expression of transcriptional isoforms Biopolym. Cell 2004 20, N 6 P. 515–523.
[3] Pucharcos C., Casas C., Nadal M., Estivill X., de la Luna S. The human intersectin genes and their spliced variants are differentially expressed Biochim. Biophys. Acta 2001 1521, N 1–3 P. 1–11.
[4] Tsyba L., Gryaznova T., Dergai O., Dergai M., Skrypkina I., Kropyvko S., Boldyryev O., Nikolaienko O., Novokhatska O., Rynditch A. Alternative splicing affecting the SH3A domain controls the binding properties of intersectin 1 in neurons Biochem. Biophys. Res. Communs 2008 372, N 4 P. 929–934.
[5] Kropyvko S., Gerasymchuk D., Skrypkina I., Dergai M., Dergai O., Nikolaienko O., Rynditch A., Tsyba L. Structural diversity and differential expression of novel human intersectin 1 isoforms Mol. Biol. Rep 2009 37, 6, pp 2789-2796
[6] Singer G. A. C., Wu J., Yan P., Plass C., Huang T. H. M., Davuluri R. V. Genome-wide analysis of alternative promoters of human genes using a custom promoter tiling array BMC Genomics 2008 9, N 349 P. 1–15.
[7] Yamashita R., Suzuki Y., Wakaguri H., Tsuritani K., Nakai K., Sugano S. DBTSS: DataBase of Human Transcription Start Sites, progress report 2006 Nucl. Acids Res 2006 34 P. 86–89.
[8] Yamabhai M., Hoffman N. G., Hardison N. L., McPherson P. S., Castagnoli L., Cesareni G., Kay B. K. Intersectin, a novel adaptor protein with two Eps15 homology and five Src homology 3 domains J. Biol. Chem 1998 273, N 47 P. 31401–31407.
[9] Tong X. K., Hussain N. K., de Heuvel E., Kurakin A., AbiJaoude E., Quinn C. C., Olson M. F., Marais R., Baranes D., Kay B. K., McPherson P. S. The endocytic protein intersectin is a major binding partner for the Ras exchange factor mSos1 in rat brain EMBO J 2000 19, N 6 P. 1263–1271.
[10] He G., Wang H., Huang S., Huang C. Intersectin links WNK kinases to endocytosis of ROMK1 J. Clin. Invest 2007 117, N 4 P. 1078–1087.