Intramolecular control of transcriptional activity by the NK2-specific domain in NK-2 homeodomain proteins

Hirotaka Watada, Raghu Mirmira, Julie Kalamaras, Michael S. German

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

The developmentally important homeodomain transcription factors of the NK-2 class contain a highly conserved region, the NK2-specific domain (NK2-SD). The function of this domain, however, remains unknown. The primary structure of the NK2-SD suggests that it might function as an accessory DNA-binding domain or as a protein-protein interaction interface. To assess the possibility that the NK2-SD may contribute to DNA-binding specificity, we used a PCR-based approach to identify a consensus DNA-binding sequences for Nkx2.2, an NK-2 family member involved in pancreas and central nervous system development. The consensus sequence (T(C)/(T)AAGT(GG)/(AC)TT) is similar to the known binding sequences for other NK-2 homeodomain proteins, but we show that the NK2-SD does not contribute significantly to specific DNA binding to this sequence. To determine whether the NK2-SD contributes to transactivation, we used GAL4-Nkx2.2 fusion constructs to map a powerful transcriptional activation domain in the C-terminal region beyond the conserved NK2-SD. Interestingly, this C-terminal region functions as a transcriptional activator only in the absence of an intact NK2-SD. The NK2-SD also can mask transactivation from the paired homeodomain transcription factor Pax6, but it has no effect on transcription by itself. These results demonstrate that the NK2-SD functions as an intramolecular regulator of the C-terminal activation domain in Nkx2.2 and support a model in which interactions through the NK2-SD regulate the ability of NK-2-class proteins to activate specific genes during development.

Original languageEnglish (US)
Pages (from-to)9443-9448
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number17
StatePublished - Aug 15 2000
Externally publishedYes

Fingerprint

Homeodomain Proteins
Transcriptional Activation
DNA
Transcription Factors
Proteins
Aptitude
Consensus Sequence
Masks
Pancreas
Central Nervous System
Polymerase Chain Reaction
NK 2
Genes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Intramolecular control of transcriptional activity by the NK2-specific domain in NK-2 homeodomain proteins. / Watada, Hirotaka; Mirmira, Raghu; Kalamaras, Julie; German, Michael S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, No. 17, 15.08.2000, p. 9443-9448.

Research output: Contribution to journalArticle

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