Identification and expression of alternatively spliced novel isoforms of cancer associated MYD88 lacking death domain in mouse
- 80 Downloads
MYD88 is an adaptor protein known to involve in activation of NF-κB through IL-1 receptor and TLR stimulation. It consists of N-terminal death domain and C-terminal Toll/IL-R homology domain that mediates its interaction with IL-1R associated kinase and IL-1R/TLR, respectively. MYD88 contributes to various types of carcinogenesis due to its involvement in oncogene induced inflammation. In the present study, we have recognized two new alternatively spliced variants of MyD88 gene in mouse using bioinformatics tools and molecular biology techniques in combination. The newly identified non-coding exon (NE-1) from 5′ upstream region alternatively splices with either exon E-2 or exon E-5 to produce two novel transcript variants MyD88N1 and MyD88N2 respectively. The transcript variant MyD88N1 was expressed in several tissues studied while the variant MyD88N2 was found to be expressed only in the brain. The analysis of the upstream region of novel exon by in silico approach revealed new promoter region PN, which possess potential signature sequences for diverse transcription factors, suggesting complex gene regulation. Studies of post translational modifications of conceptualized amino acid sequences of these isoforms revealed diversity in properties. Western blot analysis further confirmed the expression of protein isoform MYD88N1.
KeywordsMyD88 gene Alternative splicing Differential expression MYD88 isoforms Transcriptional variants
Authors are thankful to Department of Biotechnology (DBT), New Delhi, India, for generous funding to MT (Grant No. BT/PR5271/BID/7/395/2012) and for the award of Senior Research Fellowship to HMI by council of scientific and industrial research (CSIR) (Sanction No-09/112(0493)/2013-EMR-I). We are also thankful to the Department of Biochemistry A.M.U., Aligarh for providing us the necessary facilities. The funders had no role in the study design, decision to publish, or preparation of the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest in this work.
Mice (A/J) were purchased from animal house facility of Jamia Hamdard University, New Delhi, India. To bred animal in house, the institutional animal care and use committee and guidelines of the committee were followed. Experimentations were permitted by Ministry of Environment and Forests, GOI, under registration no. 714/02/a/CPCSEA. It was approved by the Institutional Animal Ethic Committee (IAEC) of Department of Biochemistry, Faculty of Life Sciences, AMU, Aligarh, India. All surgeries were performed under chloroform anaesthesia and maximum efforts were made to minimize the sufferings.
- 21.Artimo P, Jonnalagedda M, Arnold K, Baratin D, Csardi G, de Castro E, Duvaud S, Flegel V, Fortier A, Gasteiger E, Grosdidier A, Hernandez C, Ioannidis V, Kuznetsov D, Liechti R, Moretti S, Mostaguir K, Redaschi N, Rossier G, Xenarios I, Stockinger H (2012) ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res 40:597–603CrossRefGoogle Scholar
- 25.Hornbeck PV, Kornhauser JM, Tkachev S, Zhang B, Skrzypek E, Murray B, Latham V, Sullivan M (2012) PhosphoSitePlus: a comprehensive resource for investigating the structure and function of experimentally determined post-translational modifications in man and mouse. Nucleic Acids Res 40:D261–D270CrossRefGoogle Scholar
- 35.Jedrzejewski PT, Girod A, Tholey A, Konig N, Thullner S, Kinzel V, Bossemeyer D (1998) A conserved deamidation site at Asn 2 in the catalytic subunit of mammalian cAMP-dependent protein kinase detected by capillary LC-MS and tandem mass spectrometry. Protein Sci 7:457–469CrossRefPubMedPubMedCentralGoogle Scholar