3560-3562 Nucleic Acids Research, 1994, Vol. 22, No. 17
The androgen receptor
gene
Q-D" 1994 Oxford University Press
mutations database
Mark N.Patterson*, leuan A.Hughes, Bruce Gottlieb1 and Leonard Pinsky1 Department of Paediatrics, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QQ, UK and 'Lady Davis Institute for Medical Research, McGill University, 3755 Chemin de la Cote-Ste-Catherine, Montreal, Quebec H3T 1E2, Canada
ABSTRACT The androgen receptor gene mutations database is a comprehensive listing of mutations published in journals and meetings proceedings. The majority of mutations are point mutations identified in patients with androgen insensitivity syndrome. Information is included regarding the phenotype, the nature and location of the mutations, as well as the effects of the mutations on the androgen binding activity of the receptor. The current version of the database contains 149 entries, of which 114 are unique mutations. The database is available from EMBL (
[email protected]) or as a Macintosh Filemaker file (mc33001 @musica.mcgill.ca). Mutations in the androgen receptor gene give rise to defective male sexual differentiation and are associated with phenotypes covering a broad range of undervirilisation collectively referred to as androgen insensitivity syndrome (AIS) (1). Affected individuals have a 46XY karyotype, and testes which are able to synthesize and metabolize androgens normally. The syndrome is classified clinically into two forms: individuals with the complete androgen insensitivity syndrome (CAIS) have a normal female external appearance and the testes are present internally; partial androgen insensitivity syndrome (PAIS) is associated with a range of clinical presentations of ambiguous or abnormal genitalia, which may be predominantly male or female in appearance. Most of the patients in the database are classified either as CAIS or PAIS, although unusual aspects of the clinical phenotype such as the breast cancer in patients with mutations at amino acids 607 and 608 are noted in the phenotype field. The androgen receptor is a typical steroid receptor and functions as a ligand activated transcription factor (2). The receptor has three distinct functional domains and mutations are classified by which of the domains they affect. The gene encoding the receptor comprises 8 exons: exon 1 encodes the N-terminal half of the protein which is involved in transcriptional activation (3, 4); exons 2 and 3 encode the highly conserved DNA binding domain of the receptor comprising two zinc binding motifs; exons 4-8 encode the androgen binding domain. The androgen binding domain is also likely to be involved in other functions including interaction with other proteins such as hsp90 and ligand dependent transcriptional activation. The gene is located on the X chromosome, band q12, consistent with the X-linked pattern of inheritance seen in AIS (5). *To whom correspondence should be addressed
Table 1 represents a sample of the androgen receptor gene mutations database. Only published mutations are included in the database, although some have only been reported in meetings abstracts. Mutations are categorized into deletion, insertion, splice site and point mutation. There is one patient reported to contain two point mutations (6). Different laboratories employ alternative conventions for numbering nucleotides and amino acids in the androgen receptor gene/protein. The reason is that the androgen receptor gene has been cloned by several laboratories (7-10) and there are two regions within the coding sequence which are polymorphic in the normal population (11, 12). Both polymorphisms are in exon 1 and affect trinucleotide repeats: a CAG (polyglutamine) repeat ranges from 11-31 repeats (13) in the normal population, but is expanded two or three times in the adult onset form of motorneuron disease known as Kennedy's disease or spinal and bulbar muscular atrophy (14); a GGN (polyglycine) is reported to range from 16 to 24 repeats (12), although we have found alleles from 12-31(Patterson, unpublished). For the mutation database, the numbering system employed by Lubahn et al. (15) has been used, and all amino acid and nucleotide locations have therefore been converted to this convention. Using this convention the length of the CAG repeat is 20 (this gives 21 glutamines since there is a CAA codon at the 3' end of the repeat), and the GGN repeat is 24. The overall length of the androgen receptor is 919 amino acids. One of the most frequently used assays to -measure androgen receptor function is the androgen binding assay, and androgen binding parameters are therefore given where known: binding status is classified as negative, positive or defective, a term used to indicate a qulatitative or quantitative abnormality in androgen binding; Bmax (receptor concentration) and Kd (dissociation constant) are indicated as low, normal or high. The comments field is used to describe other aspects of the functional consequences of the mutations, and/or whether the mutation was identified in an unrelated patient in the same study. If the same mutation has been reported by different laboratories for unrelated patients (there is currently no system in place to indicate whether the same patient has been studied and reported by different laboratories), a separate entry is made for each patient. In this version of the database there are 149 entries representing 152 AIS patients and there are 107 different AIS mutations. The most common mutation is arg840his, caused by a CpG to TpG change, and this has been found in 6 patients. Overall, apprioximately 25 % of the point mutations involve CpG to TpG
Nucleic Acids Research, 1994, Vol. 22, No. 17 3561 Table 1. Sample from the androgen receptor gene mutations database Phenotype
Mutation Exon type
Nuc2
CAIS CAIS/MR CAIS CAIS
Deletion Deletion Deletion Deletion
1 1-8 4-8 5
743
CAIS CAIS
Insertion
1
Splice
exon4/ intron4 3
from
to
Nuc3
Nuc3
A
0
Amino from acid2 Amino acid
Domain4
to
Binding Bmax Kd
Comments
Ref
status5
Amino acid N All LBD LBD
neg neg neg neg
N LBD
neg neg
Single nucleotide deletion (18) (19) (20) Affected aunt deleted for (21) exons 6 and 7
Point PAIS/ breast cancer PAIS/ Point breast cancer CAIS Point CAIS Point PAIS Point Prostate cancer Point CAIS Point CAIS Point PAIS Point
202 gGt
gTt
2182
cGa
cAa
607
Arg
Gln
DBD
3
2185
aGg
aAg
608
Arg
Lys
DBD
pos
4 4 4
2353 2392 2469
aTt
aAt
cTg tAg
cCg tGg
664 677 703
Ile Leu Ser
Asn Pro
LBD LBD LBD
neg neg def
5 5 5 5
2550 2556 2590 2650
cGt cGa
cAt cAa
730 732 743 763
Asp Gly Tyr
LBD LBD LBD LBD
def neg def
Four nucleotide insertion
Cryptic splice-loss
(18) (22)
of amino acids 683-723
gGg
gTg
tAc
tGc
Val
Gly Met Asn Val
Cys
(23) norm
low
norm
(24)
high
(25) (26) (27)
high norm
high
(28) (29) (30) PolyGln repeat also short (31) (12 repeats). Thermolabile.
CAIS CAIS CAIS LNCaP
Point Point Point Point
6 7 7 8
2683 2887 2958 2991
cGc aTt cGt cAc
cAc aCt cAt cGc
774 842 866 877
Arg Ile Val Thr
His Thr Met Ala
LBD LBD LBD LBD
def
high
def def
norm
high
Thermolabile Altered binding
(32) (33) (34) (35)
specificity CAIS PAIS
Point Point
8 8
3081 3099
cCt gCc
cTt gTc
907 913
Leu Pro
Phe Ser
LBD LBD
def
low
norm
(36) (37)
'MR (mental retardation); LNCaP (lymph node carcinoma of the prostate cell line).
2Amino acid and nucleotide (nuc) positions are given according to the sequence of Lubahn et al (15).
3For point mutations altered bases are indicated in upper case, and unchanged bases on either side are in lower case. This is done to indicate which mutations may have arisen by deamination of a methylcytosine, but it should be noted that although three bases are indicated, they do not necessarily correspond to the codon which is changed. 4N (amino terminal 538 amino acids); DBD (DNA binding domain, amino acids 539-627); LBD (ligand binding domain, amino acids 628-919). 5Neg (negative); pos (positive and normal); def (defective in terms of qualitative or quantitative binding)
changes. The majority of the mutations are point mutations affecting the DNA or ligand binding domains. Relatively few mutations have been found in exon 1, and there are no examples of mutations causing amino acid substitutions in this exon. Alterations in this part of the receptor may therefore cause a phenotype which is somewhat different from the typical AIS phenotypes. Within the ligand binding domain, there is some evidence for clustering of amino acid substitutions. Between amino acids 664 and 913, there are 65 different amino acid substitution mutations, associated with AIS phenotypes. Clustering of the mutations occurs particularly between amino acids 741 and 774. This interval represents 13 % of the domain but contains 32 % of the mutations. As has been noted previously (16) this interval is also homologous to a region containing a cluster of mutations in the thyroid receptor beta gene, detected in patients with resistance to thyroid hormone (17). The sensitivity of this region to mutation suggests that it is critically involved in the interaction between receptor and ligand, for many members of the nuclear receptor superfamily. This database is available via anonymous FTP at FTP.EMBLHeidelberg.De in the directory pub/databases/armut. For further information send a message to the address
[email protected] containing the line HELP armut. The database
can also be obtained as a Macintosh FileMaker file from Bruce Gottlieb (
[email protected]).
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