Mutations of the mouse mu H chain which prevent polymer assembly.
Earlier work has shown that truncated mu-chains lacking the carboxy-terminal C mu 4-tail region are secreted as monomeric rather than polymeric IgM and that the monomer phenotype is not due to the lack of a disulfide bond at Cys-575 in the tail. In order to define with greater precision, the molecular requirements for IgM polymer assembly, we have isolated several mutant hybridomas which produce monomeric IgM. For three such mutants, we synthesized cDNA clones of their mu mRNA and identified a mutation in the mu-chain which was responsible for the failure to assemble polymers. Mutant 205 has a 2-bp deletion which results in a termination codon after amino acid 556, effectively deleting the last 20 amino acids of the mu-chain. In conjunction with earlier reports, this result shows that the tail plays some role in assembly other than providing Cys-575, the penultimate amino acid, for disulfide bond formation. Both mutant 21 and mutant 201 have an A to G transition, which results in Tyr-455 in the fourth constant domain being replaced by a cysteine. We conclude that the integrity of both the C mu 4 domain and the 19 amino acid tail are required for the mu H chain to be assembled into polymeric IgM.
Davis
A C
AC
Department of Immunology and Medical Genetics, University of Toronto, Canada.
Collins
C
C
Yoshimura
M I
MI
D'Agostaro
G
G
Shulman
M J
MJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005803:Genes, Immunoglobulin; D006825:Hybridomas / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007075:Immunoglobulin M / Q000096:biosynthesis* / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics*; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation
D007143:Immunoglobulin Heavy Chains; D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains; D046911:Macromolecular Substances