title: The molecular cell death machinery in the simple cnidarian Hydra includes an expanded caspase family and pro- and anti-apoptotic Bcl-2 proteins.
abstract: The fresh water polyp Hydra belongs to the phylum Cnidaria, which diverged from the metazoan lineage before the appearance of bilaterians. In order to understand the evolution of apoptosis in metazoans, we have begun to elucidate the molecular cell death machinery in this model organism. Based on ESTs and the whole Hydra genome assembly, we have identified 15 caspases. We show that one is activated during apoptosis, four have characteristics of initiator caspases with N-terminal DED, CARD or DD domain and two undergo autoprocessing in vitro. In addition, we describe seven Bcl-2-like and two Bak-like proteins. For most of the Bcl-2 family proteins, we have observed mitochondrial localization. When expressed in mammalian cells, HyBak-like 1 and 2 strongly induced apoptosis. Six of the Bcl-2 family members inhibited apoptosis induced by camptothecin in mammalian cells with HyBcl-2-like 4 showing an especially strong protective effect. This protein also interacted with HyBak-like 1 in a yeast two-hybrid assay. Mutation of the conserved leucine in its BH3 domain abolished both the interaction with HyBak-like 1 and the anti-apoptotic effect. Moreover, we describe novel Hydra BH-3-only proteins. One of these interacted with Bcl-2-like 4 and induced apoptosis in mammalian cells. Our data indicate that the evolution of a complex network for cell death regulation arose at the earliest and simplest level of multicellular organization, where it exhibited a substantially higher level of complexity than in the protostome model organisms Caenorhabditis and Drosophila.
authors.affiliation: Department Biology II, Ludwig-Maximilians University München, Planegg-Martinsried, Germany.
authors.forename: Margherita
authors.initials: M
authors.lastname: Lasi
authors.forename: Barbara
authors.initials: B
authors.lastname: Pauly
authors.forename: Nikola
authors.initials: N
authors.lastname: Schmidt
authors.forename: Mihai
authors.initials: M
authors.lastname: Cikala
authors.forename: Beate
authors.initials: B
authors.lastname: Stiening
authors.forename: Tina
authors.initials: T
authors.lastname: Käsbauer
authors.forename: Gerhardt
authors.initials: G
authors.lastname: Zenner
authors.forename: Tanja
authors.initials: T
authors.lastname: Popp
authors.forename: Anita
authors.initials: A
authors.lastname: Wagner
authors.forename: Regina T
authors.initials: RT
authors.lastname: Knapp
authors.forename: Andreas H
authors.initials: AH
authors.lastname: Huber
authors.forename: Michaela
authors.initials: M
authors.lastname: Grunert
authors.forename: Johannes
authors.initials: J
authors.lastname: Söding
authors.forename: Charles N
authors.initials: CN
authors.lastname: David
authors.forename: Angelika
authors.initials: A
authors.lastname: Böttger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000235:genetics* / Q000378:metabolism; D006829:Hydra / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D020798:Two-Hybrid System Techniques; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000235:genetics*
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D050998:bcl-2 Homologous Antagonist-Killer Protein; D020169:Caspases