eif2α kinases regulate development through the bzpr transcription factor in dictyostelium discoideumeif2α激酶调节发展dictyostelium discoideum bzpr转录因子.pdfVIP

eIF2a Kinases Regulate Development through the BzpR Transcription Factor in Dictyostelium discoideum Charles K. Singleton*, Yanhua Xiong, Janet H. Kirsten, Kelsey P. Pendleton Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee, United States of America Abstract Background: A major mechanism of translational regulation in response to a variety of stresses is mediated by phosphorylation of eIF2a to reduce delivery of initiator tRNAs to scanning ribosomes. For some mRNAs, often encoding a bZIP transcription factor, eIF2a phosphorylation leads to enhanced translation due to delayed reinitiation at upstream open reading frames. Dictyostelium cells possess at least three eIF2a kinases that regulate various portions of the starvation- induced developmental program. Cells possessing an eIF2a that cannot be phosphorylated (BS167) show abnormalities in growth and development. We sought to identify a bZIP protein in Dictyostelium whose production is controlled by the eIF2a regulatory system. Principal Findings: Cells disrupted in the bzpR gene had similar developmental defects as BS167 cells, including small entities, stalk defects, and reduced spore viability. b-galactosidase production was used to examine translation from mRNA containing the bzpR 59 UTR. While protein production was readily apparent and regulated temporally and spatially in wild type cells, essentially no b-galactosidase was produced in developing BS167 cells even though the lacZ mRNA levels were the same as those in wild type cells. Also, no protein production was observed in strains lacking IfkA or IfkB eIF2a kinases. GFP fusions, with appropriate internal controls, were used to directly demonstrate that the bzpR 59 UTR, possessing 7 uORFs, suppressed translation by 12 fold. Suppression occurred even when