Translational control of globin chain ontogeny in hamster yolk sac erythroid cells.
Prior research has demonstrated that globin ontogeny of hamster proceeds nearly to completion during the several days that yolk sac erythroid cells (YSEC) circulate in the embryo; synthesis of embryonic globin chains gives way to synthesis of adult globin chains in these primitive cells. In the present study, we translated total cell RNA extracted from YSEC on days 9-13 of gestation in wheat germ cell-free extract, expecting to observe the same progressive rise that occurs in vivo in rates of translation of alpha- and beta-globin mRNA during ontogeny. The opposite occurred; translation rates of both globins decreased sharply. This disparity between synthesis of alpha- and beta-globins in vivo and in vitro suggested an element of control of translation attributable to the YSEC cytoplasm. We therefore assayed the effect of RNA-free clarified YSEC cytoplasm on cell-free translation of YSEC RNA. A repression of translation was detected of alpha- and beta-globin mRNA (not of embryonic globin mRNA), exercised strongly by cytoplasm from YSEC early in ontogeny (gestational day 9), and weakening as ontogeny progressed. The same effect was noted on alpha- and beta-globin mRNA of adult hamster and of rabbit. Heat treatment of cytoplasm abolished the greater part of the translation regulation, suggesting that the active agent is protein. Further characterization of this translational regulator included: (a) it binds to globin poly(A) mRNA but not to poly(A), (b) it was not detected in cell lysate of adult hamster brain, lung, or erythrocytes, and (c) it did not inhibit translation of adult hamster brain and liver RNA. We conclude that hamster globin ontogeny is substantially modulated by this translational regulation of alpha- and beta-globin expression.
Lee
K M
KM
Hematology-Oncology Division, St. Luke's-Roosevelt Hospital Center, New York, New York 10025.
Bertles
J F
JF
Boussios
T
T
D000818:Animals; D002474:Cell-Free System; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005914:Globins / Q000235:genetics*; D006358:Hot Temperature; D011247:Pregnancy; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D011817:Rabbits; D013154:Spleen / Q000166:cytology / Q000378:metabolism*; D015017:Yolk Sac / Q000737:chemistry*
D012313:RNA; D005914:Globins