title: A regulatory domain in the N terminus of tryptophan hydroxylase 2 controls enzyme expression.
abstract: Serotonin is involved in a variety of physiological processes in the central nervous system and the periphery. As the rate-limiting enzyme in serotonin synthesis, tryptophan hydroxylase plays an important role in modulating these processes. Of the two variants of tryptophan hydroxylase, tryptophan hydroxylase 2 (TPH2) is expressed predominantly in the central nervous system, whereas tryptophan hydroxylase 1 (TPH1) is expressed mostly in peripheral tissues. Although the two enzymes share considerable sequence homology, the regulatory domain of TPH2 contains an additional 41 amino acids at the N terminus that TPH1 lacks. Here we show that the extended TPH2 N-terminal domain contains a unique sequence involved in the regulation of enzyme expression. When expressed in cultured mammalian cells, TPH2 is synthesized less efficiently and is also less stable than TPH1. Removal of the unique portion of the N terminus of TPH2 results in expression of the enzyme at a level similar to that of TPH1, whereas protein chimeras containing this fragment are expressed at lower levels than their wild-type counterparts. We identify a region centered on amino acids 10-20 that mediates the bulk of this effect. We also demonstrate that phosphorylation of serine 19, a protein kinase A consensus site located in this N-terminal domain, results in increased TPH2 stability and consequent increases in enzyme output in cell culture systems. Because this domain is unique to TPH2, these data provide evidence for selective regulation of brain serotonin synthesis.
authors.affiliation: Department of Neurobiology and Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Murphy
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhang
authors.forename: Raul R
authors.initials: RR
authors.lastname: Gainetdinov
authors.forename: Jean-Martin
authors.initials: JM
authors.lastname: Beaulieu
authors.forename: Marc G
authors.initials: MG
authors.lastname: Caron
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017124:Conserved Sequence; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008969:Molecular Sequence Data; D010768:Phosphoserine / Q000378:metabolism; D051381:Rats; D016415:Sequence Alignment; D012701:Serotonin / Q000096:biosynthesis; D014365:Tryptophan Hydroxylase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D010768:Phosphoserine; D012701:Serotonin; C497467:TPH1 protein, human; C494784:TPH2 protein, human; D014365:Tryptophan Hydroxylase