Purification of a glucose-binding protein from rat liver nuclei. Evidence for a role in targeting of nuclear mRNP to nuclear pore complex.
A nuclear carbohydrate-binding protein with a molecular mass of 67 kDa (CBP67), which is specific for glucose residues, was purified to essential homogeneity from rat liver nuclear extracts. This protein could also be isolated from nuclear ribonucleoprotein (RNP) complexes by extraction in the presence of 0.6 M or 2 M NaCl, but it was absent in polysomal RNP complex. The binding of the purified protein, which has an isoelectric point of 7.3, to glucose-containing glycoconjugates depends on the presence of Ca2+ and Mg2+. Using closed nuclear envelope vesicles as a system to study nuclear transport of RNA, it was shown that both entrapped polysomal mRNA and nuclear RNA precursors are readily exported from the vesicles in an ATP-dependent manner. The transport was unidirectional and strongly promoted by the poly(A) segment attached to these RNAs. In contrast, nuclear RNP complexes entrapped into the vesicles together with glucose-conjugated bovine serum albumin or nucleoplasmin, or bird nest glycoprotein, were not exported into the extravesicular space. However, transport of nuclear RNP complexes could be achieved in the presence of glucose or after co-addition of a glucose-recognizing lectin from Pellina semitubulosa. In Western blots, radioiodinated CBP67 binds to an 80-kDa polypeptide both in isolated rat liver nuclear envelopes and pore-complex laminae. From these results we postulate that CBP67 may direct nuclear RNP complexes to the nuclear pore.
Schröder
H C
HC
Institut für Physiologische Chemie, Universität Mainz, Federal Republic of Germany.
Facy
P
P
Monsigny
M
M
Pfeifer
K
K
Bek
A
A
Müller
W E
WE
D000818:Animals; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D002467:Cell Nucleus / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D007525:Isoelectric Focusing; D008099:Liver / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012261:Ribonucleoproteins / Q000378:metabolism*
D002413:Cations, Divalent; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D008274:Magnesium; D002118:Calcium