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Antibodies specific for N 6 ‐methyladenosine react with intact snRNPs U2 and U4/U6
Author(s) -
Bringmann Peter,
Lührmann Reinhard
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)81512-0
Subject(s) - snrnp , rna , ribonucleoprotein , biology , microbiology and biotechnology , biophysics , biochemistry , gene
Antibodies specific for N 6 ‐methyladenosine (m 6 A) were elicited in rabbits and used to study the accessibility in intact snRNPs of the m 6 A residues present in the snRNAs U2, U4 and U6. The antibody quantitatively precipitates snRNPs U2 and U4/U6 from total nucleoplasmic snRNPs U1–U6 isolated from HeLa cells, which demonstrates that the m 6 A residues of the respective snRNAs are not protected by snRNP proteins in the snRNP particles. While the anti‐m 6 A IgG does not react at all with U5 RNPs lacking m 6 A, a significant amount of U1 RNPs was co‐precipitated despite the fact that U1 RNA does not contain m 6 A either. Since anti‐m 6 A IgG does not react with purified U1 RNPs and co‐precipitation of U1 RNPs is dependent on the presence of U2 RNPs but not of U4/U6 RNPs, these data indicate an interaction between snRNPs U1 and U2 in vitro. The anti‐m 6 A precipitation pattern described above was also observed with snRNPs isolation from mouse Ehrlich ascites tumor cells, indicating similar three‐dimensional arrangements of snRNAs in homologous snRNP particles from different organisms.