Showing papers by "Axel Ullrich published in 1984"

Isolation of the human insulin-like growth factor I gene using a single synthetic DNA probe.

Abstract: A single synthetic oligonucleotide was employed as hybridization probe to detect and enable isolation of the human insulin-like growth factor I (IGF-I) gene from a human genomic DNA library. The synthetic oligonucleotide probe coded for the B-chain of IGF-I and was designed for expression in Escherichia coli. Despite numerous interspersed mismatches, the synthetic probe hybridized specifically with seven recombinant lambda phage containing almost the entire B-chain region of the human IGF-I gene. The usefulness of this approach was further demonstrated by the detection of lambda phage containing human preproinsulin, using A and B chain synthetic oligonucleotides, 90 and 63 nucleotides in length, as hybridization probes. The nucleotide sequence of the human IGF-I exon suggests that IGF-I is synthesized as a larger precursor molecule.

Mouse 7S nerve growth factor: complete sequence of a cDNA coding for the alpha-subunit precursor and its relationship to serine proteases.

Abstract: Two synthetic oligonucleotides, one 14-mer and one 15-mer, each containing 32 sequences and corresponding to two regions of the partially determined protein sequence, were utilized to identify three cDNA clones coding for the precursor of the alpha-subunit of 7S mouse nerve growth factor (NGF). This library, containing 860 clones, had been preselected from a much larger one by low-stringency hybridization using a cDNA probe corresponding to one of the large family of glandular kallikreins expressed in the adult male mouse submandibular gland. Partial sequence analysis had previously established the alpha-subunit to be a member of this group, albeit with no demonstrable catalytic activity. Nucleotide sequence analysis of the longest of these clones (2A4) predicted the apparent complete amino acid sequence of the 265-residue precursor. One of the other clones (3F2) contained an A----G substitution at position 565 resulting in a Lys----Glu change at position 160 of the mature sequence. These clones probably represent two different alleles. Several amino acid changes, relative to other serine proteases, are evident, which may account for the apparent lack of enzymatic activity. An Arg----Gln substitution at residue -1 would prevent cleavage of the putative activation peptide, and the deletion of residues 2-5 interrupts the highly conserved Ile/Val-Ile/Val-Gly-Gly N-terminal sequence. An Asp----Tyr substitution in the binding pocket and a Gly----His substitution near the active site serine also probably contribute to the inactive structure. The role of this subunit in NGF function remains obscure.

Structural gene for beta-nerve growth factor not defective in familial dysautonomia

Abstract: The developmental loss of neurons in sympathetic, sensory, and some parasympathetic ganglia in familial dysautonomia suggests an inherited defect in the action of beta-nerve growth factor (beta-NGF). The role of this growth factor in dysautonomia has been difficult to resolve as there is no known source of authentic human beta-NGF. The availability of a cloned DNA probe for the human beta-NGF gene has allowed identification of some copies of the gene (alleles) in six affected families. Alleles differ in the length of restriction endonuclease fragments that hybridize to DNA probes for the gene. In two families, affected children did not inherit the same two alleles at the beta-NGF locus. Since this disease is transmitted in an autosomal recessive manner, affected children must share the same alleles at the locus causing the disease. This analysis excludes the beta-NGF gene region as the cause of this neurologic disease but does not eliminate other genes involved in beta-NGF action, such as those coding for processing enzymes, receptors, or other subunits of the NGF complex.