Possible involvement of (2'5')oligoadenylate synthetase activity in pre-mRNA splicing.
The first step in splicing of pre-mRNA involves an intermediate lariat structure, in which a 2'5' phosphodiester bond between the 5' terminal guanosine residue of the intron and a specific adenosine residue near the 3' end of the intron is formed. A mammalian enzyme that generates 2'-5' phosphodiester bonds is (2'-5')oligoadenylate synthetase [(2'-5')OASE]. Although the expression of this enzyme is induced by interferon, low constitutive levels can be detected in untreated cells and tissues. The structural similarity between the lariat branch point and the 2'-5' phosphodiester bond generated by (2'-5')OASE prompted the experiments described here which suggest that this enzyme is involved in pre-mRNA splicing. (i) We show that a (2'-5')OASE activity is associated with 60S spliceosomes in an ATP- and RNA-dependent manner and that it can be indirectly immunoprecipitated by anti-Sm antibodies. (ii) Antibodies against (2'-5')OASE inhibit the lariat formation in the first step of splicing when added directly to a splicing reaction in vitro. (iii) HeLa cell nuclear extracts immunodepleted of (2'-5')OASE activity were also deficient in splicing activity.
Sperling
J
J
Department of Organic Chemistry, Weizmann Institute of Science, Rehovot, Israel.
Chebath
J
J
Arad-Dann
H
H
Offen
D
D
Spann
P
P
Lehrer
R
R
Goldblatt
D
D
Jolles
B
B
Sperling
R
R
D015088:2',5'-Oligoadenylate Synthetase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D002467:Cell Nucleus / Q000201:enzymology*; D005914:Globins / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000494:pharmacology; D007700:Kinetics; D008970:Molecular Weight; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D012326:RNA Splicing; D011817:Rabbits; D012261:Ribonucleoproteins / Q000276:immunology / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear
D000906:Antibodies; D012322:RNA Precursors; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D005914:Globins; D007372:Interferons; D015088:2',5'-Oligoadenylate Synthetase