Dokument

Zusammenfassung:
Die Koordination von Wachstum und Differenzierung als Antwort auf verschiedene Signale und Umweltbedingungen ist eine grundlegende Eigenschaft von Organismen. Hierbei erfordern die Anpassung von Zellteilungszyklus und Stoffwechsel und morphologische Änderungen eine spezifische Regulation der Genexpression mit Hilfe von Transkriptionsfaktoren. Eine zentrale Frage ist, welche Kombinationen von Transkriptionsfaktoren an welchen Promotoren bei der Regulation von Wachstum und Differenzierung überhaupt eine Rolle spielen. In der vorliegenden Arbeit wurde exemplarisch die kombinatorische und promotorspezifische Kontrolle des TEA-Transkriptionsfaktors Tec1 aus der Bäckerhefe Saccharomyces cerevisiae untersucht. Tec1 ist zusammen mit dem Transkriptionsfaktor Ste12 an der Regulation der Differenzierungsprogramme Biofilmbildung und Konjugation beteiligt und wird unter anderem über den Fus3/Kss1-MAPK-Signalweg gesteuert. Im ersten Teil dieser Arbeit wurde die Komplexbildung von Tec1 und Ste12 analysiert. Es stellte sich heraus, dass Tec1-Zielgene in eine Ste12-abhängig und in eine Ste12-unabhängig regulierte Klasse eingeteilt werden können. Ste12 kann die Stabilität von Tec1 über dessen C-terminale Domäne regulieren, die zudem eine Ste12-unabhängige Transkriptionsaktivierungsdomäne enthält. Im zweiten Teil der Arbeit wurden die Co-Regulatoren Msa1 und Msa2 als neue Interaktionspartner von Tec1 identifiziert. Tec1, Msa1 und Msa2 regulieren gemeinsam zelluläre Differenzierungsprogramme wie Biofilmbildung und Konjugation. Zudem wirken sie auf den Zellteilungszyklus und die Ribosomensynthese. Dies sind Prozesse, die das Wachstum und die Zellgröße beeinflussen. Tec1, Ste12, Msa1 und Msa2 können verschiedene Komplexe ausbilden und in unterschiedlichen Kombinationen an gemeinsam regulierte Promotoren binden. Der dritte Teil dieser Arbeit beschreibt den Aufbau und die Validierung eines Mess-systems, mit dessen Hilfe man in lebenden Zellen die Signaltransduktionsdynamik im Fus3/Kss1-MAPK-Modul unter verschiedenen Bedingungen quantitativ messen kann. Dies geschieht durch Bestimmung von Menge, Lokalisierung und programm-spezifischer Aktivität der Regulatoren Tec1, Ste12, Dig1 und Dig2 mittels Fluoreszenz-mikroskopie.

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