Apolipoprotein B mRNA and lipoprotein secretion are increased in McArdle RH-7777 cells by expression of betaine-homocysteine S-methyltransferase.
The cDNA encoding rat betaine-homocysteine S-methyltransferase (BHMT) was isolated through production of monoclonal antibodies against protein fractions enriched with apolipoprotein B (apo B)-mRNA-editing complexes. BHMT mRNA was expressed predominantly in liver, and also in kidney, but not in small intestine. In stable McArdle RH-7777 (McA) cell lines expressing differing levels of BHMT, the editing efficiency of apo B mRNA was unchanged. Evaluation of apo B-mRNA expression revealed that steady-state levels were increased significantly and in parallel with BHMT protein expression. The highest levels of BHMT mRNA and BHMT enzyme activity expressed in stably transfected McA cells were comparable with those found in rat hepatocytes. In contrast to the changes in apo B-mRNA abundance, levels of other apolipoprotein-encoding mRNAs and several liver-specific and ubiquitously expressed mRNAs were unchanged by BHMT expression. In the cell line expressing the highest level of BHMT, apo B-containing lipoprotein secretion was increased, indicating utilization of increased endogenous message. Results suggest that apo B-mRNA abundance in McA cells is related to the expression of BHMT, an enzyme important in homocysteine metabolism.
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, 601 Elmwood Avenue, Rochester, NY 14642, USA.
M P
MP
Sowden
H L
HL
Collins
H C
HC
Smith
T A
TA
Garrow
J D
JD
Sparks
C E
CE
Sparks
D000595:Amino Acid Sequence; D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D050940:Betaine-Homocysteine S-Methyltransferase; D018076:DNA, Complementary; D007422:Intestines / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008780:Methyltransferases / Q000235:genetics*; D008969:Molecular Sequence Data; D017393:RNA Editing; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; D018076:DNA, Complementary; D012333:RNA, Messenger; D008780:Methyltransferases; D050940:Betaine-Homocysteine S-Methyltransferase; C494550:Bhmt protein, rat