TY - JOUR
T1 - On the existence of polyadenylated histone mRNA in Xenopus laevis oocytes
AU - Levenson, Robert
AU - Marcu, Kenneth B.
N1 - Funding Information:
This work was carried out in part by R . G . L . in the laboratory of Dr. R . W . Merriam in partial fulfillment of the requirements for the Ph .D . degree . We wish to thank Drs . Joseph Kates for oligo(dT)-cellulose ; Robert P . Perry for the use of his oligo(dT)-cellulose column ; Martin Nemer for L . pictus and S . purpuratus embryos ; and John Brooker for wheat germ(A)+ RNA . We also thank Dennis O'Kane for his excellent technical assistance in carrying out tryptic digest experiments . We especially wish to thank Drs. Robert Perry, Martin Nemer, and Bernard Dudock for making available to us the use of equipment and providing laboratory space for parts of this project . Finally, we wish to thank Drs . R . P . Perry, M . Nemer, B . Dudock, D . Poccia, R . W . Merriam, and J . Kates for invaluable discussions and help during the course of this research, and Drs . R . P . Perry, D . Housman, and D . Poccia for critical readings of the manuscript prior to publication . This work was supported by a Stony Brook Biomedical Sciences Support grant to R . W . Mer-riam ; by grants from the NIH and the American Cancer Society to B. Dudock; by a grant from the NSF to Robert P . Perry; and by an appropriation from the Commonwealth of Pennsylvania . K.B.M . was supported by an NIH postdoctoral fellowship.
PY - 1976/10
Y1 - 1976/10
N2 - In a variety of systems, histone mRNA has been shown to lack poly(A) (Adesnik and Darnell, 1972;Grunstein et al., 1973). We have found, however, that in Xenopus laevis oocytes, poly(A)-containing mRNA codes for histones, in a wheat germ cell-free system, based on the following criteria: first, co-migration with authentic X. laevis oocyte histones on polyacrylamide gels; second, no detectable incorporation of tryptophan; third, differential incorporation of lysine and methionine into histone fraction H2A; fourth, resistance of histone fraction H2A to cleavage with cyanogen bromide; and fifth, correspondence of tryptic peptide maps of partially purified cell-free products with authentic X. laevis oocyte histone. RNA which directs the synthesis of histones in the cell-free system is retained on oligo(dT)-cellulose, even after denaturation in 80% DMSO at 70°C, thereby demonstrating the covalent attachment of polyadenylic acid sequences to the mRNA. Poly(A)- RNA (7S-14S fraction) was also found to code for histones using the same criteria. We discuss the significance of the finding that X. laevis oocytes contain two classes of histone mRNA as well as the potential developmental implications of this observation.
AB - In a variety of systems, histone mRNA has been shown to lack poly(A) (Adesnik and Darnell, 1972;Grunstein et al., 1973). We have found, however, that in Xenopus laevis oocytes, poly(A)-containing mRNA codes for histones, in a wheat germ cell-free system, based on the following criteria: first, co-migration with authentic X. laevis oocyte histones on polyacrylamide gels; second, no detectable incorporation of tryptophan; third, differential incorporation of lysine and methionine into histone fraction H2A; fourth, resistance of histone fraction H2A to cleavage with cyanogen bromide; and fifth, correspondence of tryptic peptide maps of partially purified cell-free products with authentic X. laevis oocyte histone. RNA which directs the synthesis of histones in the cell-free system is retained on oligo(dT)-cellulose, even after denaturation in 80% DMSO at 70°C, thereby demonstrating the covalent attachment of polyadenylic acid sequences to the mRNA. Poly(A)- RNA (7S-14S fraction) was also found to code for histones using the same criteria. We discuss the significance of the finding that X. laevis oocytes contain two classes of histone mRNA as well as the potential developmental implications of this observation.
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U2 - 10.1016/0092-8674(76)90121-5
DO - 10.1016/0092-8674(76)90121-5
M3 - Article
C2 - 987857
AN - SCOPUS:0017103802
VL - 9
SP - 311
EP - 322
JO - Cell
JF - Cell
SN - 0092-8674
IS - 2
ER -