Changing patterns in cytoskeletal mRNA expression and protein synthesis during murine erythropoiesis in vivo.
The major cytoskeletal proteins alpha-spectrin, beta-spectrin, and ankyrin are synthesized and assembled into a supportive membrane skeleton during erythroid differentiation. Information on the temporal appearance of mRNA and protein species is essential for understanding both the cytoskeletal assembly process and the function of various isoforms. We have isolated highly enriched populations of fetal erythroid cells at various stages of maturation. mRNAs for erythroid ankyrin, alpha-spectrin, and beta-spectrin were expressed at all stages but there were differences in transcript types and levels. The ratio of 9-kilobase (kb) to 7.5-kb erythroid ankyrin transcripts decreased markedly during differentiation, but there was no change in the ratio of the 10.1-kb and 9.3-kb erythroid beta-spectrin transcripts. The relative amounts of ankyrin, alpha-spectrin, and beta-spectrin mRNA increased during yolk sac cell differentiation, whereas only alpha-spectrin mRNA increased during differentiation of the fetal liver cells. The amounts of beta-spectrin mRNA exceeded the amounts of alpha-spectrin mRNA in the early precursors from both yolk sac and fetal liver; protein synthetic levels showed the same pattern. The 16-day fetal peripheral reticulocytes, on the other hand, had the adult mRNA and protein synthetic ratios with alpha/beta greater than 1. The data indicate that at least two mechanisms exist to meet changing erythroid membrane cytoskeletal requirements during development in utero: (i) stage-specific processing of the mRNA for the major cytoskeletal linker protein ankyrin and (ii) developmentally regulated alpha/beta-spectrin protein synthetic rates.
Peters
L L
LL
Jackson Laboratory, Bar Harbor, ME 04609.
White
R A
RA
Birkenmeier
C S
CS
Bloom
M L
ML
Lux
S E
SE
Barker
J E
JE
D000818:Animals; D017487:Ankyrins; D001798:Blood Proteins / Q000235:genetics; D003599:Cytoskeleton / Q000378:metabolism*; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D004920:Erythropoiesis; D015870:Gene Expression; D008099:Liver / Q000196:embryology; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010659:Phenylhydrazines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D013049:Spectrin / Q000235:genetics / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism; D015017:Yolk Sac
D017487:Ankyrins; D001798:Blood Proteins; D008565:Membrane Proteins; D010659:Phenylhydrazines; D012333:RNA, Messenger; C030299:phenylhydrazine; D013049:Spectrin