Polymorphisms in the 3′-untranslated region of human and monkey dopamine transporter genes affect reporter gene expression

被引:199
作者
Miller, GM [1 ]
Madras, BK [1 ]
机构
[1] Harvard Univ, Sch Med, New England Reg Primate Res Ctr, Div Neurochem, Southborough, MA 01772 USA
关键词
luciferase; cocaine; VNTR; ADHD; SNP; HSV-TK; SV40; cloning; promoter; interaction;
D O I
10.1038/sj/mp/4000921
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Dopamine transporter (DAT) levels vary in normal subjects and deviate from the normal range in pathological states. We investigated mechanisms by which the DAT gene may influence DAT protein expression. As the 3'-untransiated region (3'-UTR) of the DAT gene varies with regard to length and single nucleotide pollymorphisms (SNPs), we addressed whether the 3'UTR of sequence-defined DAT alleles can differentially affect the level of reporter gene expression in vitro. We first established that within individual rhesus monkeys, two alleles of the DAT gene were expressed in the substantia nigra. We then transfected HEK-293 cells with HSV-TK- and SV40-driven luciferase expression vectors harboring downstream DAT 3'-UTR segments of alleles containing polymorphisms of length (human: 9 or 10 repeat units) or SNPs within alleles of fixed length (human: Dral-sensitive (Dral+) vs Dral-insensitive (Dral-) 10-repeat alleles; rhesus monkey: Bst1107l-sensitive (Bst+) vs Bst1107l-insensitive (Bst-) 12-repeat alleles). Vectors containing the 3'-UTR segment of a human DAT allele containing nine tandem repeat units resulted in significantly higher levels of luciferase production than analogous vectors containing 10 tandem repeat units. Depending on the promoter used, vectors containing the human or monkey 3'-UTR segments that differed on the basis of an SNP resulted in increases or decreases in luciferase gene expression. This report provides experimental evidence that variability in the length or the sequence of the 3'-UTR of the DAT gene may influence levels of DAT protein in the brain.
引用
收藏
页码:44 / 55
页数:12
相关论文
共 85 条
[1]   Tenotomy decreases reporter protein synthesis via the 3′-untranslated region of the β-myosin heavy chain mRNA [J].
Ashley, WW ;
Russell, B .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2000, 279 (01) :C257-C265
[2]   DOPAMINE TRANSPORTER MESSENGER-RNA CONTENT IN HUMAN SUBSTANTIA-NIGRA DECREASES PRECIPITOUSLY WITH AGE [J].
BANNON, MJ ;
POOSCH, MS ;
YUE, X ;
GOEBEL, DJ ;
CASSIN, B ;
KAPATOS, G .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (15) :7095-7099
[3]   Biogenic amine transporters: regulation in flux [J].
Blakely, RD ;
Bauman, AL .
CURRENT OPINION IN NEUROBIOLOGY, 2000, 10 (03) :328-336
[4]   Plasma homovanillic acid and the dopamine transporter during cocaine withdrawal [J].
Bowers, MB ;
Malison, RT ;
Seibyl, JP ;
Kosten, TR .
BIOLOGICAL PSYCHIATRY, 1998, 43 (04) :278-281
[5]  
CHAN PT, 1990, J BIOL CHEM, V265, P4091
[6]   Comparison of the role of dopamine, serotonin, and noradrenaline genes in ADHD, ODD and conduct disorder: multivariate regression analysis of 20 genes [J].
Comings, DE ;
Gade-Andavolu, R ;
Gonzalez, N ;
Wu, SJ ;
Muhleman, D ;
Blake, H ;
Dietz, G ;
Saucier, G ;
MacMurray, JP .
CLINICAL GENETICS, 2000, 57 (03) :178-196
[7]  
Comings DE, 1996, AM J MED GENET, V67, P264, DOI 10.1002/(SICI)1096-8628(19960531)67:3<264::AID-AJMG4>3.0.CO
[8]  
2-N
[9]   Multivariate analysis of associations of 42 genes in ADHD, ODD and conduct disorder [J].
Comings, DE ;
Gade-Andavolu, R ;
Gonzalez, N ;
Wu, SJ ;
Muhleman, D ;
Blake, H ;
Chiu, F ;
Wang, E ;
Farwell, K ;
Darakjy, S ;
Baker, R ;
Dietz, G ;
Saucier, G ;
MacMurray, P .
CLINICAL GENETICS, 2000, 58 (01) :31-40
[10]  
COOK EH, 1995, AM J HUM GENET, V56, P993