Molecular cloning and characterization of bovine beta-mannosidase.
Deficiency of lysosomal beta-mannosidase activity results in a severe neurodegenerative disease in goats and cattle and a relatively milder phenotype in humans. A cDNA coding for the entire beta-mannosidase protein is described. Mixed oligonucleotides derived from bovine beta-mannosidase peptide sequences were used to screen a bovine thyroid cDNA library. Clones covering about 80% of the C-terminal region were recovered. The missing 5'-region was obtained using the technique of 5'-rapid amplification of cDNA ends. The composite cDNA contains 3852 nucleotides, encoding 879 amino acids. The N-terminal methionine is followed by 16 amino acids displaying the characteristics of a typical signal peptide sequence. The deduced amino acid sequence is colinear with all peptide sequences determined by protein microsequencing. Northern blot analysis demonstrates a single 4.2-kilobase transcript in various tissues from both normal and affected goats and calves. The mRNA level is decreased in tissues of affected beta-mannosidosis animals. The gene encoding beta-mannosidase is localized to human chromosome 4 as shown by Southern analysis of rodent/human somatic cell hybrids. This is the first report of cloning of lysosomal beta-mannosidase.
Chen
H
H
Department of Pathology, Michigan State University, East Lansing 48824.
Leipprandt
J R
JR
Traviss
C E
CE
Sopher
B L
BL
Jones
M Z
MZ
Cavanagh
K T
KT
Friderici
K H
KH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008247:Lysosomes / Q000201:enzymology; D008361:Mannosidases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D044902:beta-Mannosidase
D018076:DNA, Complementary; D008361:Mannosidases; D044902:beta-Mannosidase