RESUMO
Amylase expression in strain YBR differs in several respects from the standard mouse phenotype. The synthesis of salivary amylase is elevated twofold in YBR mice and the synthesis of pancreatic amylase is reduced to one-half the normal rate. We have compared the concentrations of amylase mRNA in the parotid, liver and pancreas of YBR mice with those in strains A/J and C3H. We observed differences in amylase mRNA abundance which can account for the levels of amylase protein synthesis in the parotid and pancreas of these strains. Unexpectedly, the concentration of amylase mRNA in the liver of YBR mice was also higher than in the other strains. Since liver amylase is transcribed from the same gene as parotid amylase, duplication of the Amy-1 locus could account for the elevated mRNA concentration in both tissues. Quantitative analysis of genomic DNA by Southern blotting provided direct evidence for duplication of Amy-1 in strain YBR.
Assuntos
Genes , Variação Genética , Fígado/enzimologia , Pâncreas/enzimologia , Glândula Parótida/enzimologia , RNA Mensageiro/genética , alfa-Amilases/genética , Animais , Cinética , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos , Peso Molecular , Hibridização de Ácido Nucleico , Especificidade de Órgãos , Especificidade da EspécieRESUMO
The two isozymes of pancreatic amylase in mouse strain YBR/Ki are encoded by closely linked genes which are independently regulated. We have isolated these two pancreatic amylase genes, Amy-2.1 and Amy-2.2, from a cosmid library of YBR/Ki genomic DNA and compared the nucleotide sequences of coding regions with the amino acid sequences of the protein isozymes. Transcripts of both genes were also isolated from a pancreatic cDNA library and partially sequenced. The results demonstrate that Amy-2.1 encodes the A1 isozyme of YBR/Ki pancreatic amylase, while Amy- 2.2 encodes the insulin-dependent B1 isozyme. Similarities of restriction maps and nucleotide sequences suggest that Amy-2.1 is closely related to the active Amy-2a gene previously isolated from strain A/J (Schibler, U., Pittet, A.-C., Young, R. A., Hagenbüchle, O., Tosi, M., Gellman, S., and Wellauer, P. K. (1982) J. Mol. Biol. 155, 247-266). Expression of Amy-2.2 may be limited to strain YBR/Ki. The inactive Amy-X gene from A/J (Schibler, U., Pittet, A.-C., Young, R. A., Hagenbüchle, O., Tosi, M., Gellman, S., and Wellauer, P. K. (1982) J. Mol. Biol. 155, 247-266) is apparently a null allele of Amy-2.2. An additional amylase gene from YBR/Ki has been identified as a pancreatic amylase pseudogene which diverged between sixteen and thirty-two million years ago. The pancreatic amylase subfamily in strain YBR/Ki thus consists of two active genes and one pseudogene. The low rate of amylase production in YBR/Ki pancreas, relative to that of other inbred strains, can be accounted for by the lower number of gene copies in this strain. Comparison of pancreatic amylase genes from different inbred strains provides evidence for several duplication and deletion events during the recent evolution of this chromosome region.
Assuntos
Amilases/genética , Pâncreas/enzimologia , Animais , Sequência de Bases , Evolução Biológica , Mapeamento Cromossômico , DNA/isolamento & purificação , Isoenzimas/genética , Camundongos , Camundongos Endogâmicos , RNA Mensageiro/metabolismo , Especificidade da Espécie , Transcrição GênicaRESUMO
The var1 locus of yeast mitochondrial DNA encodes a protein of the small mitochondrial ribosome subunit, denoted var1 protein. The size of var1 protein was previously shown to exhibit a strain-dependent polymorphism, determined by various combinations of at least three genetic elements. We report here that the var1 gene is itself polymorphic and that the six forms of this gene examined here differ by various combinations of three in-frame insertions into the coding region of the smallest allele. These insertions, which appear to be the molecular basis for the genetic elements, could increase the size of var1 protein by 8, 10, 16, 24, or 26 amino acid residues, accounting for the observed protein polymorphisms. Furthermore, we have characterized three additional sources of sequence variation located outside of the coding region but within major transcripts of the var1 gene.