title: Unconventional translation initiation of human trypsinogen 4 at a CUG codon with an N-terminal leucine. A possible means to regulate gene expression.
abstract: Chromosomal rearrangements apparently account for the presence of a primate-specific gene (protease serine 3) in chromosome 9. This gene encodes, as the result of alternative splicing, both mesotrypsinogen and trypsinogen 4. Whereas mesotrypsinogen is known to be a pancreatic protease, neither the chemical nature nor biological function of trypsinogen 4 has been explored previously. The trypsinogen 4 sequence contains two predicted translation initiation sites: an AUG site that codes for a 72-residue leader peptide on Isoform A, and a CUG site that codes for a 28-residue leader peptide on Isoform B. We report studies that provide evidence for the N-terminal amino acid sequence of trypsinogen 4 and the possible mechanism of expression of this protein in human brain and transiently transfected cells. We raised mAbs against a 28-amino acid synthetic peptide representing the leader sequence of Isoform B and against recombinant trypsin 4. By using these antibodies, we isolated and chemically identified trypsinogen 4 from extracts of both post mortem human brain and transiently transfected HeLa cells. Our results show that Isoform B, with a leucine N terminus, is the predominant (if not exclusive) form of the enzyme in post mortem human brain, but that both isoforms are expressed in transiently transfected cells. On the basis of our studies on the expression of a series of trypsinogen 4 constructs in two different cell lines, we propose that unconventional translation initiation at a CUG with a leucine, rather than a methionine, N terminus may serve as a means to regulate protein expression.
authors.affiliation: Department of Biochemistry, Eötvös Loránd University, Pázmány Peter s. 1/C, Budapest H-1117, Hungary.
authors.forename: Attila L
authors.initials: AL
authors.lastname: Németh
authors.forename: Péter
authors.initials: P
authors.lastname: Medveczky
authors.forename: Júlia
authors.initials: J
authors.lastname: Tóth
authors.forename: Erika
authors.initials: E
authors.lastname: Siklódi
authors.forename: Katalin
authors.initials: K
authors.lastname: Schlett
authors.forename: András
authors.initials: A
authors.lastname: Patthy
authors.forename: Miklós
authors.initials: M
authors.lastname: Palkovits
authors.forename: Judit
authors.initials: J
authors.lastname: Ovádi
authors.forename: Natália
authors.initials: N
authors.lastname: Tõkési
authors.forename: Péter
authors.initials: P
authors.lastname: Németh
authors.forename: László
authors.initials: L
authors.lastname: Szilágyi
authors.forename: László
authors.initials: L
authors.lastname: Gráf
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D003062:Codon; D004247:DNA; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D007930:Leucine / Q000737:chemistry*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D014362:Trypsinogen / Q000737:chemistry / Q000235:genetics*
chemical_list: D003062:Codon; D012333:RNA, Messenger; D014362:Trypsinogen; D004247:DNA; D007930:Leucine