Deletion of etoposide-induced 2.4 kb transcript (ei24) reduced cell proliferation and aggregate-size in Dictyostelium discoideum

作者：Gupta Neha; Saran Shweta^*

来源：International Journal of Developmental Biology, 2018, 62(4-5): 273-283.

DOI：10.1387/ijdb.170327ss

• Dictyostelium EI24
• cAMP signalling
• etoposide
• DNA-damage
• cell cycle

The etoposide-induced 2.4 kb transcript (ei24)gene is induced both by p53 and etoposide, an anti-cancer tumour drug. There is no p53 gene present in Dictyostelium discoideum. Thus, the functions of ei24 in the absence of p53 were analysed. Both overexpressor (ei24(OE)) and knockout (ei24(-)) mutants were made to study its role during growth, development and differentiation. Additionally, cell cycle and its response to DNA-damage were also analysed.We identified, characterized and elucidated the functions of the ei24 gene in Dictyostelium. In silico analyses demonstrated the conservation across eukaryotes and in situ hybridization showed it to be prestalk-specific. ei24(-) cells showed reduced cell proliferation and cell-cohesive properties, ultimately forming small-sized aggregates that developed into miniature and stalky fruiting bodies. The ei24(OE) cells formed fruiting bodies with engorged or double-decker type son with short stalks.The ei24(-) cells showed reduced cAMP signalling with lower intracellular cAMP levels resulting in diminished migration of cells along cAMP gradients. Deletion of ei24 resulted in mis-expression of prestalk-specific markers. Cell cycle analysis revealed an increased bias towards the stalk-pathway by ei24(-) cells and vice-versa for ei24(O)(E) cells. EI24 in Dietyostelium functions even in the absence of p53 and is induced in response to both UV-radiation and etoposide treatments. ei24(OE) cells showed enhanced DNA-damage repair mechanisms. Also, etoposide treatment and overexpression of ei24 caused G2/M arrest in the cell cycle. Our results indicate that EI24 is important for the growth, development and differentiation of Dietyostelium apart from being a DNA-damage response gene.

• 出版日期2018