title: Evidence that two Pcl-like cyclins control Cdk9 activity during cell differentiation in Aspergillus nidulans asexual development.
abstract: Cyclin-dependent protein kinases (CDKs) are usually involved in cell cycle regulation. However, Cdk9 is an exception and promotes RNA synthesis through phosphorylation of the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAPII). The CTD is comprised of repeating heptapeptides, in which serine residues at positions 2, 5, and 7 are of crucial importance. Ser5 phosphorylation causes transcription initiation and promoter escape. However, RNAPII pauses 20 to 50 bp downstream from the transcription start site, until Cdk9 phosphorylates Ser2. This event relieves the checkpoint and promotes the processivity of elongation. Here we present evidence that in the filamentous fungus Aspergillus nidulans, a Cdk9 homologue, PtkA, serves specific functions in conidiophore development. It was previously shown that PtkA interacts with two cyclins, PclA and the T cyclin PchA. Using yeast two-hybrid screens, we identified a third cyclin, PclB, and a kinase, PipA(Bud32). Both proteins were expressed in hyphae and in conidiophores, but interaction between each protein and PtkA was restricted to the conidiophores. Deletion of pchA caused a severe growth defect, and deletion of pipA was lethal, suggesting basic functions in PtkA-dependent gene transcription. In contrast, deletion of pclB in combination with deletion of pclA essentially caused a block in spore formation. We present evidence that the phosphorylation status of the CTD of RNA polymerase II in the conidiophore changes upon deletion of pclA or pclB. Our results suggest that tissue-specific modulation of Cdk9 activity by PclA and PclB is required for proper differentiation.
authors.affiliation: Karlsruhe Institute of Technology, Institute for Applied Biosciences, Department of Microbiology, Karlsruhe, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Kempf
authors.forename: Friederike
authors.initials: F
authors.lastname: Bathe
authors.forename: Reinhard
authors.initials: R
authors.lastname: Fischer
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000201:enzymology / Q000254:growth & development / Q000502:physiology*; D002467:Cell Nucleus / Q000201:enzymology; D002468:Cell Physiological Phenomena; D017124:Conserved Sequence; D042863:Cyclin-Dependent Kinase 9 / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D010766:Phosphorylation; D011485:Protein Binding; D060066:Protein Interaction Maps; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012319:RNA Polymerase II / Q000378:metabolism; D012100:Reproduction, Asexual; D012441:Saccharomyces cerevisiae; D013172:Spores, Fungal / Q000201:enzymology / Q000254:growth & development / Q000502:physiology
chemical_list: D005656:Fungal Proteins; C429306:PclA protein, Aspergillus; D042863:Cyclin-Dependent Kinase 9; D012319:RNA Polymerase II