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Identification and expression of alternatively spliced novel isoforms of cancer associated MYD88 lacking death domain in mouse

  • Hassan Mubarak Ishqi
  • Mohammed Amir Husain
  • Sayeed Ur Rehman
  • Tarique Sarwar
  • Mohammad Tabish
Original Article
  • 56 Downloads

Abstract

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.

Keywords

MyD88 gene Alternative splicing Differential expression MYD88 isoforms Transcriptional variants 

Notes

Acknowledgements

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.

Ethical approval

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.

Supplementary material

11033_2018_4209_MOESM1_ESM.tif (5.8 mb)
Supplementary Figure 1 Multiple sequence alignment (Clustal Omega) of amino acid sequence of published and new isoforms of MYD88 in mouse. The names of the isoforms are shown adjacent to the sequence. The alignment clearly depicts the marked difference at the N-termini of these isoforms. The MYD88N1 sequence starts from the methionine present in exon E-3 which is represented by downward red arrow. The translation initiation point in case of MYD88N2 is present in the exon E-5 shown with the downward green arrow. The asterisk (*) and dash (-) shows identical residues and no residues respectively (TIF 5974 KB)
11033_2018_4209_MOESM2_ESM.tif (3.7 mb)
Supplementary Figure 2 CpG Plot generated by EMBOSS Cpgplot tool used for CpG Island prediction. The predicted promoter region sequence was used for all the promoters. CpG Islands were present in the regions of promoter PC and PN with observed/expected ratio > 0.60 and G + C percentage > 60. However, the putative CpG Islands were found in PN only (TIF 3744 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Biochemistry, Faculty of Life SciencesA.M. UniversityAligarhIndia
  2. 2.Department of BiosciencesJamia Millia IslamiaNew DelhiIndia
  3. 3.Deutsches Diabetes-ZentrumLeibniz-Zentrum für Diabetes-Forschung an der Heinrich-Heine-Universität DüsseldorfDüsseldorfGermany

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