F-actin homeostasis through transcriptional regulation and proteasome-mediated proteolysis

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 115; no. 28; pp. E6487 - E6496
• Main Authors: Onishi, Masayuki, Pecani, Kresti, Jones, Taylor, Pringle, John R., Cross, Frederick R.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 10.07.2018
• Series: PNAS Plus
• Subjects: Actin; Actins - biosynthesis; Actins - genetics; Biological Sciences; Cell division; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - genetics; Chlamydomonas reinhardtii - metabolism; Cleavage; Cofilin; Cytoskeleton; Deactivation; Degradation; Feedback loops; Forming; Gene regulation; Gene sequencing; Genes; Growth conditions; Homeostasis; latrunculin B; Plant Proteins - genetics; Plant Proteins - metabolism; PNAS Plus; Profilin; Proteasome Endopeptidase Complex - genetics; Proteasome Endopeptidase Complex - metabolism; Proteasomes; Proteins; Proteolysis; Ribonucleic acid; RNA; Transcription; Transcription, Genetic; Ubiquitin; proteasome; actin; algal cytoskeletons; latrunculin; Chlamydomonas
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1721935115

Many organisms possess multiple and often divergent actins whose regulation and roles are not understood in detail. For example, Chlamydomonas reinhardtii has both a conventional actin (IDA5) and a highly divergent one (NAP1); only IDA5 is expressed in normal proliferating cells. We showed previously that the drug latrunculin B (LatB) causes loss of filamentous (F-) IDA5 and strong up-regulation of NAP1, which then provides essential actin function(s) by forming LatB-resistant F-NAP1. RNA-sequencing analyses now show that this up-regulation of NAP1 reflects a broad transcriptional response, much of which depends on three proteins (LAT1, LAT2, and LAT3) identified previously as essential for NAP1 transcription. Many of the LAT-regulated genes contain a putative cis-acting regulatory site, the “LRE motif.” The LatB transcriptional program appears to be activated by loss of F-IDA5 and deactivated by formation of F-NAP1, thus forming an F-actin–dependent negative-feedback loop. Multiple genes encoding proteins of the ubiquitin-proteasome system are among those induced by LatB, resulting in rapid degradation of IDA5 (but not NAP1). Our results suggest that IDA5 degradation is functionally important because nonpolymerizable LatB-bound IDA5 interferes with the formation of F-NAP1. The genes for the actin-interacting proteins cofilin and profilin are also induced. Cofilin induction may further the clearance of IDA5 by promoting the scission of F-IDA5, whereas profilin appears to function in protecting monomeric IDA5 from degradation. This multifaceted regulatory system allows rapid and quantitative turnover of F-actin in response to cytoskeletal perturbations and probably also maintains F-actin homeostasis under normal growth conditions.