{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T05:21:17Z","timestamp":1791264077803,"version":"4.1.0"},"reference-count":55,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5355","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1998,2,27]]},"abstract":"<jats:p>\n            The nocturnal increase in circulating melatonin in vertebrates is regulated by 10- to 100-fold increases in pineal serotonin\n            <jats:italic>N<\/jats:italic>\n            -acetyltransferase (AA-NAT) activity. Changes in the amount of AA-NAT protein were shown to parallel changes in AA-NAT activity. When neural stimulation was switched off by either light exposure or\n            <jats:sc>l<\/jats:sc>\n            -propranolol\u2013induced \u03b2-adrenergic blockade, both AA-NAT activity and protein decreased rapidly. Effects of\n            <jats:sc>l<\/jats:sc>\n            -propranolol were blocked in vitro by dibutyryl adenosine 3\u2032,5\u2032-monophosphate (cAMP) or inhibitors of proteasomal proteolysis. This result indicates that adrenergic-cAMP regulation of AA-NAT is mediated by rapid reversible control of selective proteasomal proteolysis. Similar proteasome-based mechanisms may function widely as selective molecular switches in vertebrate neural systems.\n          <\/jats:p>","DOI":"10.1126\/science.279.5355.1358","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:35:26Z","timestamp":1027762526000},"page":"1358-1360","source":"Crossref","is-referenced-by-count":237,"title":["Melatonin Production: Proteasomal Proteolysis in Serotonin\n            <i>N<\/i>\n            -Acetyltransferase Regulation"],"prefix":"10.1126","volume":"279","author":[{"given":"Jonathan A.","family":"Gastel","sequence":"first","affiliation":[{"name":"Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, MD 20892\u20134480, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick H.","family":"Roseboom","sequence":"additional","affiliation":[{"name":"Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, MD 20892\u20134480, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter A.","family":"Rinaldi","sequence":"additional","affiliation":[{"name":"Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, MD 20892\u20134480, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joan L.","family":"Weller","sequence":"additional","affiliation":[{"name":"Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, MD 20892\u20134480, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David C.","family":"Klein","sequence":"additional","affiliation":[{"name":"Section on Neuroendocrinology, Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development (NICHD), National Institutes of Health, Bethesda, MD 20892\u20134480, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"crossref","first-page":"1992","DOI":"10.1016\/S0021-9258(18)69351-2","volume":"235","author":"Lerner A. B.","year":"1960","unstructured":"Lerner A. B., Case J. D., Takahashi Y., J. Biol. Chem. 235, 1992 (1960);","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_2_3","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1126\/science.184.4144.1341","volume":"184","author":"Axelrod J.","year":"1974","unstructured":"Axelrod J., Science 184, 1341 (1974);","journal-title":"Science"},{"key":"e_1_3_1_2_4","unstructured":"; J. Arendt Melatonin and the Mammalian Pineal Gland (Chapman & Hall London 1995) pp. 1\u2013273; F. J. Karsch C. J. I. Woodfill B. Malpaux J. E. Robinson N. L. Wayne in The Suprachiasmatic Nucleus: The Mind's Clock D. C. Klein R. Y. Moore S. M. Reppert Eds. (Oxford Univ. Press New York 1991) pp. 217\u2013230."},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.169.3950.1093"},{"key":"e_1_3_1_3_3","doi-asserted-by":"crossref","unstructured":"; ibid. 177 532 (1972);","DOI":"10.1126\/science.177.4048.532"},{"key":"e_1_3_1_3_4","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1016\/0024-3205(71)90360-2","volume":"10","author":"Illnerova H.","year":"1971","unstructured":"Illnerova H., Life Sci. 10, 955 (1971);","journal-title":"Life Sci."},{"key":"e_1_3_1_3_5","doi-asserted-by":"crossref","first-page":"848","DOI":"10.1210\/endo-96-4-848","volume":"96","author":"Binkley S.","year":"1975","unstructured":"Binkley S., Macbride S. E., Klein D. C., Ralph C. L., Endocrinology 96, 848 (1975);","journal-title":"Endocrinology"},{"key":"e_1_3_1_3_6","first-page":"482","volume":"98","author":"Rollag M. D.","year":"1976","unstructured":"Rollag M. D., Niswender G. D., ibid. 98, 482 (1976).","journal-title":"ibid."},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.167.3926.1738"},{"key":"e_1_3_1_4_3","first-page":"163","volume":"184","author":"Brownstein M.","year":"1974","unstructured":"Brownstein M., Axelrod J., ibid. 184, 163 (1974);","journal-title":"ibid."},{"key":"e_1_3_1_4_4","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1042\/bst0200299","volume":"20","author":"Klein D. C.","year":"1992","unstructured":"Klein D. C., Schaad N., Namboodiri M. A. A., Yu L., Weller J. L., Biochem. Soc. Trans. 20, 299 (1992).","journal-title":"Biochem. Soc. Trans."},{"key":"e_1_3_1_5_2","unstructured":"Rat AA-NAT (rAA-NAT) antisera (As) 2500 and As2559 were raised against phospho-rAA-NAT 22\u201337 (accession number gbU38306) or a mixture of rAA-NAT 49\u201359 rAA-NAT 72\u201385 and rAA-NAT 89\u2013100 respectively (19). As2821 was raised against purified bacterially expressed rAA-NAT 50\u2013150 fused to measles fusion protein 288\u2013302 (accession number p41356) ["},{"key":"e_1_3_1_5_3","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1016\/0161-5890(92)90202-9","volume":"29","author":"Partidos C.","year":"1992","unstructured":"Partidos C., Stanley C., Steward M., Mol. Immunol. 29, 651 (1992);","journal-title":"Mol. Immunol."},{"key":"e_1_3_1_5_4","unstructured":"]. irAA-NAT on protein immunoblots (19) was quantitated with a Storm PhosphorImager (Molecular Dynamics) or from nonsaturated autoradiographs (X-O-MAT film Kodak) which were digitized (Microtek Scanmaker II; Adobe Photoshop version 3.05) and analyzed with NIH Image version 1.57 software. A unit of immunoreactive protein is approximately equal to the signal generated by \u223c30 pg of bacterially expressed rAANAT determined with As2559. This is 1% of the signal typically generated by a rat pineal gland obtained in the middle of the night [Zeitgeber time (ZT) 21]."},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1016\/S0076-6879(87)42069-7","volume":"142","author":"Namboodiri M. A. A.","year":"1986","unstructured":"Namboodiri M. A. A., Dubbels R., Klein D. C., Methods Enzymol. 142, 583 (1986).","journal-title":"Methods Enzymol."},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"2547","DOI":"10.1073\/pnas.69.9.2547","volume":"69","author":"Deguchi T.","year":"1972","unstructured":"Deguchi T., Axelrod J., Proc. Natl. Acad. Sci. U.S.A. 69, 2547 (1972).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_8_2","first-page":"2107","volume":"72","author":"Romero J. A.","year":"1975","unstructured":"Romero J. A., Zatz M., Axelrod J., ibid. 72, 2107 (1975);","journal-title":"ibid."},{"key":"e_1_3_1_8_3","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1111\/j.1471-4159.1979.tb00407.x","volume":"32","author":"Illnerova H.","year":"1979","unstructured":"Illnerova H., Vanecek J., Krecek J., Wetterberg L., S\u00e4\u00e4f J., J. Neurochem. 32, 673 (1979).","journal-title":"J. Neurochem."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.1210\/endo.137.7.8770929","volume":"137","author":"Roseboom P. H.","year":"1996","unstructured":"Roseboom P. H., et al., Endocrinology 137, 3033 (1996);","journal-title":"Endocrinology"},{"key":"e_1_3_1_9_3","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1046\/j.1471-4159.1997.68010213.x","volume":"68","author":"Bernard M.","year":"1997","unstructured":"Bernard M., et al., J. Neurochem. 68, 213 (1997);","journal-title":"J. Neurochem."},{"key":"e_1_3_1_9_4","unstructured":"; D. C. Klein et al. Recent Prog. Hormone Res. 52 307 (1997)."},{"key":"e_1_3_1_10_2","unstructured":"Norepinepherine isoproteronol dibutyryl cAMP and 8-bromocyclic AMP reproducibly increased AA-NAT activity (2) and irAA-NAT (4) in parallel in pineal organ culture (2) and in isolated pinealocytes (20)."},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.202027"},{"key":"e_1_3_1_12_2","unstructured":"Cells (\u223c10 6 ) were homogenized in 200 \u03bcl of 50 mM tris (pH 7.5) 500 mM NaCl 1 mM dithiothreitol (DTT) and 100 \u03bcM phenylmethylsulfonyl fluoride (PMSF); the homogenate was then centrifuged and AA-NAT was immunoprecipitated with 2 \u03bcl of As2821 (2 hours 4\u00b0C) and protein A\u2013Sepharose (1 hour 4\u00b0C). AA-NAT activity and irAA-NAT are stable under these conditions. AA-NAT was labeled by incubation (1 hour 30\u00b0C final volume = 20 \u03bcl) with 10 \u03bcCi of [ 32 \u03b3]ATP (adenosine triphosphate) 100 mM NaCl 50 mM tris-HCl (pH 7.5) 1 mM DTT 10 mM MgCl and 33 U of protein kinase A (PKA Promega); radioactive bands were analyzed and quantitated as described (4). This technique is quantitative within the range of AA-NAT values used in these experiments."},{"key":"e_1_3_1_13_2","unstructured":"Rat pinealocytes or pineal glands were treated in experiments similar to those in Fig. 3. Unless otherwise indicated the concentration of all protease inhibitors was 100 \u03bcM. The following protease inhibitors (targeted class) were ineffective: (serine) PMSF (Sigma) leupeptin (ICN) aprotinin (ICN) N-p -tosyl- l -phenylalanine chloromethyl ketone (Calbiochem); (lysosomal) chloroquine (0.2 mM Sigma); (aspartic) pepstatin (ICN); (metallo) ethylenediamine tetraacetic acid (Sigma); (cysteine) trans -epoxysuccinyl- l -leucylamido (4-guanidino)butane (Sigma) calpeptin (Calbiochem) and \u03b1- N -acetyl-leucine-leucine-methioninal (Calbiochem). The following protease or proteasome inhibitors preserved more than 50% of the activity or AA-NAT protein compared with drug alone: calpain inhibitor I and Mg115 ["},{"key":"e_1_3_1_13_3","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1016\/S0092-8674(94)90462-6","volume":"78","author":"Rock K. L.","year":"1994","unstructured":"Rock K. L., et al., Cell 78, 761 (1994);","journal-title":"Cell"},{"key":"e_1_3_1_13_4","unstructured":"] Mg132 (V. J. Palombella O. J. Rando A. L. Goldberg T. Maniatis ibid. p. 773) Z-leucine-leucine-leucine-vinyl sulfone ["},{"key":"e_1_3_1_13_5","doi-asserted-by":"crossref","first-page":"6629","DOI":"10.1073\/pnas.94.13.6629","volume":"94","author":"Bogyo M.","year":"1997","unstructured":"Bogyo M., et al., Proc. Natl. Acad. Sci. U.S.A. 94, 6629 (1997);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_13_6","unstructured":"] lactacystin ["},{"key":"e_1_3_1_13_7","doi-asserted-by":"publisher","DOI":"10.1126\/science.7732382"},{"key":"e_1_3_1_13_8","unstructured":"] and \u03b2-clastolactacystin ["},{"key":"e_1_3_1_13_9","doi-asserted-by":"crossref","first-page":"7273","DOI":"10.1074\/jbc.271.13.7273","volume":"271","author":"Dick L. R.","year":"1997","unstructured":"Dick L. R., Cruikshank L., Grenier F. D., Melandri S. L., Stein R. L., J. Biol. Chem. 271, 7273 (1997)].","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_14_2","unstructured":"Ubiquitination occurs via \u03b5-amino groups of lysines in target proteins and is required to target many proteins for proteasomal degradation ["},{"key":"e_1_3_1_14_3","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1146\/annurev.bi.61.070192.003553","volume":"61","author":"Hershko L. A.","year":"1992","unstructured":"Hershko L. A., Ciechanover A., Annu. Rev. Biochem. 61, 761 (1992);","journal-title":"Annu. Rev. Biochem."},{"key":"e_1_3_1_14_4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genet.30.1.405"},{"key":"e_1_3_1_14_5","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.65.070196.004101"},{"key":"e_1_3_1_14_6","unstructured":"]. An NH 2 -terminal lysine residue is present in all known vertebrate AA-NAT sequences; this region is important for regulation of AA-NAT activity and irAA-NAT by cAMP and proteosome inhibitors in C6 glioma cells transiently transfected with AA-NAT (J. A. Gastel and D. C. Klein unpublished observations)."},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5242.1681"},{"key":"e_1_3_1_16_2","unstructured":"The rapid suppressive effects of light on melatonin production are evolutionarily conserved: rat (6) human ["},{"key":"e_1_3_1_16_3","doi-asserted-by":"publisher","DOI":"10.1126\/science.7434030"},{"key":"e_1_3_1_16_4","unstructured":"] chicken ["},{"key":"e_1_3_1_16_5","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/0006-8993(83)90660-1","volume":"266","author":"Hamm H. E.","year":"1983","unstructured":"Hamm H. E., Takahashi J. S., Menaker M., Brain Res. 266, 287 (1983);","journal-title":"Brain Res."},{"key":"e_1_3_1_16_6","unstructured":"] and fish ["},{"key":"e_1_3_1_16_7","doi-asserted-by":"crossref","first-page":"1943","DOI":"10.1523\/JNEUROSCI.09-06-01943.1989","volume":"9","author":"Falcon J.","year":"1989","unstructured":"Falcon J., Marmillon J. B., Claustrat B., Collin J. P., J. Neurosci. 9, 1943 (1989)].","journal-title":"J. Neurosci."},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","first-page":"2367","DOI":"10.1172\/JCI117929","volume":"95","author":"Costelli P.","year":"1995","unstructured":"Costelli P., et al., J. Clin. Invest. 95, 2367 (1995);","journal-title":"J. Clin. Invest."},{"key":"e_1_3_1_17_3","doi-asserted-by":"crossref","first-page":"1897","DOI":"10.1056\/NEJM199612193352507","volume":"335","author":"Mitch W. E.","year":"1996","unstructured":"Mitch W. E., Goldberg A. L., N. Engl. J. Med. 335, 1897 (1996);","journal-title":"N. Engl. J. Med."},{"key":"e_1_3_1_17_4","first-page":"H329","volume":"265","author":"Decker R. S.","year":"1993","unstructured":"Decker R. S., et al., Am. J. Physiol. 265, H329 (1993).","journal-title":"Am. J. Physiol."},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"7436","DOI":"10.1073\/pnas.90.16.7436","volume":"90","author":"Hegde A. N.","year":"1993","unstructured":"Hegde A. N., Goldberg A. L., Schwartz J. H., Proc. Natl. Acad. Sci. U.S.A. 90, 7436 (1993);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_18_3","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0092-8674(00)80188-9","volume":"89","author":"Hegde A. N.","year":"1997","unstructured":"Hegde A. N., et al., Cell 89, 115 (1997).","journal-title":"Cell"},{"key":"e_1_3_1_19_2","doi-asserted-by":"crossref","first-page":"953","DOI":"10.1126\/science.563102","volume":"198","author":"Tamarkin L.","year":"1977","unstructured":"Tamarkin L., Hollister C. W., Lefebvre N. G., Goldman B. D., Science 198, 953 (1977);","journal-title":"Science"},{"key":"e_1_3_1_19_3","doi-asserted-by":"crossref","first-page":"2276","DOI":"10.1210\/endo-113-6-2276","volume":"113","author":"Bittman E. L.","year":"1983","unstructured":"Bittman E. L., Dempsey R. J., Karsch F. J., Endocrinology 113, 2276 (1983).","journal-title":"Endocrinology"},{"key":"e_1_3_1_20_2","first-page":"713","volume":"25","author":"Weller J.","year":"1996","unstructured":"Weller J., Jaffe H., Roseboom P. H., Zylka M. J., Klein D. C., Brain Res. 25, 713 (1996).","journal-title":"Brain Res."},{"key":"e_1_3_1_21_2","doi-asserted-by":"crossref","first-page":"6979","DOI":"10.1074\/jbc.272.11.6979","volume":"272","author":"Baler R.","year":"1997","unstructured":"Baler R., Covington S., Klein D. C., J. Biol. Chem. 272, 6979 (1997).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_22_2","unstructured":"D. W. Wayne Biostatistics (Wiley New York ed. 6 1995) p. 210; ibid. pp. 598\u2013599."},{"key":"e_1_3_1_23_2","unstructured":"We thank S. Coon (NICHD) and R. Baler (NICHD) for technical advice; P. Loh (NICHD) M. Bogyo and H. Ploegh (Massachusetts Institute of Technology) for helpful discussions; and A. Ciechanover A. Goldberg and M. Zatz for use of reagents and equipment. Supported by a Pharmacology Research Associate Training (PRAT) fellowship from NIGMS (J.A.G.)."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.279.5355.1358","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T05:56:59Z","timestamp":1705125419000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.279.5355.1358"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,2,27]]},"references-count":55,"journal-issue":{"issue":"5355","published-print":{"date-parts":[[1998,2,27]]}},"alternative-id":["10.1126\/science.279.5355.1358"],"URL":"https:\/\/doi.org\/10.1126\/science.279.5355.1358","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,2,27]]}}}