The human hnRNP-M proteins: structure and relation with early heat shock-induced splicing arrest and chromosome mapping.
With anti-hnRNP monoclonal antibody 6D12 we previously showed in HeLa cells that as early as 10 min after the onset of a heat shock at 45 degrees C, a 72.5-74 kDa antigen doublet leaves the hnRNPs and strongly associates with the nuclear matrix, the effect being reversed after a 6 h recovery at 37 degrees C. cDNA cloning and sequencing enabled us to identify these antigens as hnRNP-M proteins and further to show that the correct sequence differs by an 11 amino acid stretch from the originally published sequence. We also show that monoclonal antibodies raised against synthetic hnRNP-M peptides can directly inhibit in vitro splicing. Furthermore, stressing cells at 45 degrees C for 10 min is sufficient to abolish the splicing capacity of subsequently prepared nuclear extracts which, interestingly, do not contain the hnRNP-M proteins any more. Taken together, our data suggest that these proteins are involved in splicing as well as in early stress-induced splicing arrest. Further in situ hybridization assays located the hnRNP-M encoding gene on human chromosome 19.
Gattoni
R
R
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, C.U. De Strasbourg, France.
Mahé
D
D
Mähl
P
P
Fischer
N
N
Mattei
M G
MG
Stévenin
J
J
Fuchs
J P
JP
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002874:Chromosome Mapping; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D006367:HeLa Cells; D018869:Heat-Shock Response / Q000235:genetics*; D034761:Heterogeneous-Nuclear Ribonucleoprotein Group M; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D012326:RNA Splicing; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012261:Ribonucleoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D013347:Subcellular Fractions / Q000378:metabolism
D000911:Antibodies, Monoclonal; D034761:Heterogeneous-Nuclear Ribonucleoprotein Group M; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D011994:Recombinant Proteins; D012261:Ribonucleoproteins