Induction of calreticulin expression in response to amino acid deprivation in Chinese hamster ovary cells.
The role of calreticulin as a stress-induced molecular chaperone protein of the endoplasmic reticulum is becoming more apparent. We characterize here the induction of calreticulin in response to complete amino acid deprivation in Chinese hamster ovary cells. Amino acid deprivation caused a 4-fold increase in calreticulin protein levels over a period of 4-10 h. In addition to an overall increase in protein levels, the glycosylation of calreticulin was increased. This glycosylation event was blocked by tunicamycin and was not required for the increase in calreticulin protein levels. Immunofluorescence studies localized calreticulin to the ER of CHO cells, and no significant change was observed after amino acid deprivation. Northern-blot analysis showed that calreticulin mRNA levels were increased approx. 10-fold in response to complete amino acid deprivation. The response was sensitive to actinomycin D and alpha-amanitin, implying that regulation is primarily at the level of transcription. These results are similar to the large increases in asparagine synthetase mRNA observed in response to amino acid deprivation, but the amino acid-deprivation-response element identified to be involved in asparagine synthetase induction is absent from the calreticulin promoter.
Heal
R
R
Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol BS8 1TD, U.K.
McGivan
J
J
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D037282:Calreticulin; D006224:Cricetinae; D005786:Gene Expression Regulation; D006031:Glycosylation; D018384:Oxidative Stress; D012333:RNA, Messenger / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D000596:Amino Acids; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D012333:RNA, Messenger; D012261:Ribonucleoproteins