Vol. 3, Issue 1, Part A (2016)
Immune stimulation of Anopheles gambiae 4a3B cells induces chromatin reorganization at the Defensin 1 gene
Author(s): Sandra Rosas-Madrigal, Alejandra Cabrera, Verónica Valverde-Garduno
Abstract: Mosquito innate immunity is a critical component in the transmission of Plasmodium parasites that cause malaria in human populations. Molecular mechanisms involved in regulation of transcription of mosquito innate immunity genes are not yet fully understood. Here, the chromatin conformation at the Defensin 1 (Def1) gene was investigated in the Anopheles gambiae derived, immune-responsive, 4a3B cell line. Our aim was to characterize chromatin structure, before and after immune challenge, at this innate immunity gene. Here we report that, upon immune stimulation with a heat-killed bacterial challenge, Def1 promoter sequences become co-localized with an open chromatin site. This indicates that the Def1 promoter undergoes a dynamic chromatin transition upon immune challenge. In contrast, promoter-flanking sequences and exon sequences were not nucleosome depleated in the same samples, obtained from immune stimulated cell cultures. Taken together, these results suggest participation of chromatin modification in the transcriptional activation of the Def1 gene.
Open chromatin profiling protocol comparison. Diagram shows key steps in preparation of open chromatin samples. The main difference between protocols is the use of a single step for cell lysis in the fast prep protocol; detection can be done by qPCR or semi quantitative PCR. Other steps are carried out in the same way in both protocols.
How to cite this article:
Sandra Rosas-Madrigal, Alejandra Cabrera, Verónica Valverde-Garduno. Immune stimulation of Anopheles gambiae 4a3B cells induces chromatin reorganization at the Defensin 1 gene. Int J Mosq Res 2016;3(1):30-34.