| RRC ID |
37706
|
| 著者 |
Nakayama M, Kato T, Susa T, Sano A, Kitahara K, Kato Y.
|
| タイトル |
Dimeric PROP1 binding to diverse palindromic TAAT sequences promotes its transcriptional activity.
|
| ジャーナル |
Mol Cell Endocrinol
|
| Abstract |
Mutations in the Prop1 gene are responsible for murine Ames dwarfism and human combined pituitary hormone deficiency with hypogonadism. Recently, we reported that PROP1 is a possible transcription factor for gonadotropin subunit genes through plural cis-acting sites composed of AT-rich sequences containing a TAAT motif which differs from its consensus binding sequence known as PRDQ9 (TAATTGAATTA). This study aimed to verify the binding specificity and sequence of PROP1 by applying the method of SELEX (Systematic Evolution of Ligands by EXponential enrichment), EMSA (electrophoretic mobility shift assay) and transient transfection assay. SELEX, after 5, 7 and 9 generations of selection using a random sequence library, showed that nucleotides containing one or two TAAT motifs were accumulated and accounted for 98.5% at the 9th generation. Aligned sequences and EMSA demonstrated that PROP1 binds preferentially to 11 nucleotides composed of an inverted TAAT motif separated by 3 nucleotides with variation in the half site of palindromic TAAT motifs and with preferential requirement of T at the nucleotide number 5 immediately 3' to a TAAT motif. Transient transfection assay demonstrated first that dimeric binding of PROP1 to an inverted TAAT motif and its cognates resulted in transcriptional activation, whereas monomeric binding of PROP1 to a single TAAT motif and an inverted ATTA motif did not mediate activation. Thus, this study demonstrated that dimeric binding of PROP1 is able to recognize diverse palindromic TAAT sequences separated by 3 nucleotides and to exhibit its transcriptional activity.
|
| 巻・号 |
307(1-2)
|
| ページ |
36-42
|
| 公開日 |
2009-8-13
|
| DOI |
10.1016/j.mce.2009.03.010
|
| PII |
S0303-7207(09)00200-7
|
| PMID |
19524124
|
| MeSH |
Amino Acid Sequence
Animals
Base Sequence
Consensus Sequence
Electrophoretic Mobility Shift Assay
Homeodomain Proteins / chemistry
Homeodomain Proteins / genetics*
Homeodomain Proteins / metabolism*
Inverted Repeat Sequences / genetics*
Molecular Sequence Data
Protein Binding
Protein Multimerization*
Regulatory Sequences, Nucleic Acid / genetics
Sequence Alignment
Sus scrofa
Transcriptional Activation / genetics*
|
| IF |
3.871
|
| 引用数 |
7
|
|
WOS 分野
|
ENDOCRINOLOGY & METABOLISM
CELL BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
|
