Changes in heat shock protein synthesis and hsp70 gene transcription during erythropoiesis of Xenopus laevis

Abstract

As erythroid cells of Xenopus laevis differentiate, their capacity for transcription and protein synthesis becomes progressively restricted. Mature erythrocytes are thought to be transcriptionally inert and have very low levels of protein synthesis ( Maclean et al., 1973 , Cell Differ. 2, 261–269). We have examined actual and potential synthetic activity during erythropoiesis by comparing the patterns of protein synthesis and measuring the transcriptional activity of selected genes under ambient and heat shock conditions. As erythropoiesis proceeds, there is a progressive decrease in the number of heat shock proteins (hsps) whose synthesis is induced at elevated temperatures. Despite this repression, mature erythrocytes are able to respond to heat shock with the induction of synthesis of one protein, hsp70. In addition, an hsp70-like protein is synthesized constitutively in orthochromatic erythroblasts, the final erythroid cell stage before maturation into erythrocytes. An in vitro nuclear run-off transcription assay was used to study transcription of the hsp70, β-globin, and ribosomal genes during erythropoiesis. β-Globin and ribosomal gene transcription were detected at all stages of erythropoiesis, including the erythrocyte stage. However, transcription of the β-globin gene relative to ribosomal gene transcription declines substantially in erythrocytes as compared to earlier stages. Hsp70 gene transcription shows both stage-specific and heat shock-dependent transcription. Orthochromatic erythroblasts and erythrocytes have heat shock-dependent hsp70 gene transcription, whereas the gene is transcribed at the earlier erythroblast stages at ambient temperature as well as during heat shock. The constitutive hsp70 transcripts made during these earlier stages may be stored for utilization in constitutive synthesis of the hsp70-like protein in orthochromatic erythroblasts. These data indicate that although erythrocytes become synthetically repressed during differentiation, they are less inert than previously thought. The data also reveal that the synthesis of hsps and related proteins is subject to both transcriptional and post-transcriptional control during erythropoiesis.