Ectopic expression of the atypical HLH FaPRE1 gene determines changes in cell size and morphology

Plant Science ◽  
2021 ◽  
Vol 305 ◽  
pp. 110830
Author(s):  
L. Medina-Puche ◽  
F.J. Martínez-Rivas ◽  
F.J. Molina-Hidalgo ◽  
J.A. García-Gago ◽  
J.A. Mercado ◽  
...  
2020 ◽  
Author(s):  
Shuchun Liu ◽  
Shanwen Ke ◽  
Guojian Tang ◽  
Dahui Huang ◽  
Minyi Wei ◽  
...  

Abstract Rice caryopses are enclosed by the outside glumes. It is long been acknowledged that the size and dimension of the outer glume is the main determinant that dictates the caryopsis size. However, it is unclear whether the development of caryopsis is completely dependent on the size of the glume, or it can grow and expand autonomously in spite of the constraint of glume enclosure. Here we report the identification of a mutant line that produces normal-sized glumes with smaller mature caryopsis that do not fill up the entire glume cavity. The mutant phenotype is caused by ectopic expression of a leucine-rice repeat extensin gene, OsPEX1 . The caryopsis phenotype in pex1 was caused by a reduction in cell size. OsPEX1 is highly expressed in the developing caryopsis. Over expression of the OsPEX1 gene driven by a constitutive promoter recapitulates the mutant phenotype, confirming that the small caryopsis phenotype is caused by ectopic expression of the OsPEX1 gene. Our results suggest that caryopsis development can be genetically uncoupled from maternally controlled glume development.


mBio ◽  
2020 ◽  
Vol 11 (5) ◽  
Author(s):  
Gro Anita Stamsås ◽  
Marine Restelli ◽  
Adrien Ducret ◽  
Céline Freton ◽  
Pierre Simon Garcia ◽  
...  

ABSTRACT Control of peptidoglycan assembly is critical to maintain bacterial cell size and morphology. Penicillin-binding proteins (PBPs) are crucial enzymes for the polymerization of the glycan strand and/or their cross-linking via peptide branches. Over the last few years, it has become clear that PBP activity and localization can be regulated by specific cognate regulators. The first regulator of PBP activity in Gram-positive bacteria was discovered in the human pathogen Streptococcus pneumoniae. This regulator, named CozE, controls the activity of the bifunctional PBP1a to promote cell elongation and achieve a proper cell morphology. In this work, we studied a previously undescribed CozE homolog in the pneumococcus, which we named CozEb. This protein displays the same membrane organization as CozE but is much more widely conserved among Streptococcaceae genomes. Interestingly, cozEb deletion results in cells that are smaller than their wild-type counterparts, which is the opposite effect of cozE deletion. Furthermore, double deletion of cozE and cozEb results in poor viability and exacerbated cell shape defects. Coimmunoprecipitation further showed that CozEb is part of the same complex as CozE and PBP1a. However, although we confirmed that CozE is required for septal localization of PBP1a, the absence of CozEb has no effect on PBP1a localization. Nevertheless, we found that the overexpression of CozEb can compensate for the absence of CozE in all our assays. Altogether, our results show that the interplay between PBP1a and the cell size regulators CozE and CozEb is required for the maintenance of pneumococcal cell size and shape. IMPORTANCE Penicillin-binding proteins (PBPs), the proteins catalyzing the last steps of peptidoglycan assembly, are critical for bacteria to maintain cell size, shape, and integrity. PBPs are consequently attractive targets for antibiotics. Resistance to antibiotics in Streptococcus pneumoniae (the pneumococcus) are often associated with mutations in the PBPs. In this work, we describe a new protein, CozEb, controlling the cell size of pneumococcus. CozEb is a highly conserved integral membrane protein that works together with other proteins to regulate PBPs and peptidoglycan synthesis. Deciphering the intricate mechanisms by which the pneumococcus controls peptidoglycan assembly might allow the design of innovative anti-infective strategies, for example, by resensitizing resistant strains to PBP-targeting antibiotics.


Blood ◽  
2005 ◽  
Vol 105 (11) ◽  
pp. 4477-4483 ◽  
Author(s):  
Peter S. Hammerman ◽  
Casey J. Fox ◽  
Morris J. Birnbaum ◽  
Craig B. Thompson

Abstract The Akt kinases promote hematopoietic cell growth and accumulation through phosphorylation of apoptotic effectors and stimulation of mTOR-dependent translation. In Akt-transformed leukemic cells, tumor growth can be inhibited by the mTOR inhibitor rapamycin, and clinical trials of rapamycin analogs for the treatment of leukemia are under way. Surprisingly, nontransformed hematopoietic cells can grow and proliferate in the presence of rapamycin. Here, we show that Pim-2 is required to confer rapamycin resistance. Primary hematopoietic cells from Pim-2– and Pim-1/Pim-2–deficient animals failed to accumulate and underwent apoptosis in the presence of rapamycin. Although animals deficient in Akt-1 or Pim-1/Pim-2 are viable, few animals with a compound deletion survived development, and those that were born had severe anemia. Primary hematopoietic cells from Akt-1/Pim-1/Pim-2–deficient animals displayed marked impairments in cell growth and survival. Conversely, ectopic expression of either Pim-2 or Akt-1 induced increased cell size and apoptotic resistance. However, though the effects of ectopic Akt-1 were reversed by rapamycin or a nonphosphorylatable form of 4EBP-1, those of Pim-2 were not. Coexpression of the transgenes in mice led to additive increases in cell size and survival and predisposed animals to rapid tumor formation. Together, these data indicate that Pim-2 and Akt-1 are critical components of overlapping but independent pathways, either of which is sufficient to promote the growth and survival of nontransformed hematopoietic cells.


2020 ◽  
Author(s):  
Dong Ju Lee ◽  
Il-Chul Kim ◽  
Seungwoo Jeong ◽  
Seung Gon Wi ◽  
Suk Weon Kim ◽  
...  

Abstract BackgroundThe Capsicum annuum receptor-like kinase 1 (CaRLK1) gene encodes a transmembrane protein with a cytoplasmic kinase domain and an extracellular domain. It functions as a negative regulator of plant cell death. Ectopic expression of CaRLK1 showed the hypoxia-resistance and enhanced cell division and proliferation. In this study, it was investigated which genes were controlled by ectopic expression of CaRLK1 because it decreased its average cell size of transgenic RLK1ox cells compared with that of wild-type cells (BY-2). ResultsThe average diameter (AD) of the protoplast of RLK1ox cells was about 10 µm shorter than the AD of BY-2 cells. The diameter distribution of the circular protoplasts is mainly from 20 to 40 μm (71.7%) in RLK1ox cells, whereas from 30 to 50 μm (73.0%) in BY-2 cells. Furthermore, cell volume of RLK1ox cells is also 2.5-times smaller than that of BY-2 cells. Smaller cell size of the RLK1ox cells may be related with the inhibited cell expansion because expressions of 12 expansin A genes, 6 expansin-like B genes, 2 ACC synthase genes, and 5 ACC oxidase genes were suppressed, but expressions of 7 gibberellin 2-oxidase genes were induced. The cell walls between two RLK1ox cells are approximately 138.83±4.12 nm thick on average, while those between two BY-2 cells are approximately 156.58±4.54 nm thick. Furthermore, the total content of neutral sugars in the cell wall of RLK1ox cells is less than that of BY-2 cells (about 25%). The RLK1ox cells contained 30% less glucose content than did the BY-2 cells. Thinner cell wall of the RLK1ox cells is related with the decreased cellulose biosynthesis and hemicellulose biosynthesis. Expressions of 7 CESA genes, 5 sucrose synthase 1 genes, 3 mannose-1-phosphate guanyltransferase genes, and 6 glucomannan-4-beta-mannosyltransferase genes were suppressed. The suppressed expressions of 14 polygalacturonase genes may also contribute to make the cell wall of RLK1ox cell thinner than that of BY-2 cell. ConclusionOf special emphasis is its impact of CaRLK1 gene on cell size control and cell wall thickness. Smaller cell size of the RLK1ox cells correlates with the inhibited cell expansion.


mSphere ◽  
2020 ◽  
Vol 5 (5) ◽  
Author(s):  
Cristina Velázquez-Suárez ◽  
Ignacio Luque ◽  
Antonia Herrero

ABSTRACT The model cyanobacterium Anabaena sp. PCC 7120 exhibits a phototrophic metabolism relying on oxygenic photosynthesis and a complex morphology. The organismic unit is a filament of communicated cells that may include cells specialized in different nutritional tasks, thus representing a paradigm of multicellular bacteria. In Anabaena, the inorganic carbon and nitrogen regime influenced not only growth, but also cell size, cell shape, and filament length, which also varied through the growth cycle. When using combined nitrogen, especially with abundant carbon, cells enlarged and elongated during active growth. When fixing N2, which imposed lower growth rates, shorter and smaller cells were maintained. In Anabaena, gene homologs to mreB, mreC, and mreD form an operon that was expressed at higher levels during the phase of fastest growth. In an ntcA mutant, mre transcript levels were higher than in the wild type and, consistently, cells were longer. Negative regulation by NtcA can explain that Anabaena cells were longer in the presence of combined nitrogen than in diazotrophic cultures, in which the levels of NtcA are higher. mreB, mreC, and mreD mutants could grow with combined nitrogen, but only the latter mutant could grow diazotrophically. Cells were always larger and shorter than wild-type cells, and their orientation in the filament was inverted. Consistent with increased peptidoglycan width and incorporation in the intercellular septa, filaments were longer in the mutants, suggesting a role for MreB, MreC, and MreD in the construction of septal peptidoglycan that could affect intercellular communication required for diazotrophic growth. IMPORTANCE Most studies on the determination of bacterial cell morphology have been conducted in heterotrophic organisms. Here, we present a study of how the availability of inorganic nitrogen and carbon sources influence cell size and morphology in the context of a phototrophic metabolism, as found in the multicellular cyanobacterium Anabaena. In Anabaena, the expression of the MreB, MreC, and MreD proteins, which influence cell size and length, are regulated by NtcA, a transcription factor that globally coordinates cellular responses to the C-to-N balance of the cells. Moreover, MreB, MreC, and MreD also influence septal peptidoglycan construction, thus affecting filament length and, possibly, intercellular molecular exchange that is required for diazotrophic growth. Thus, here we identified new roles for Mre proteins in relation to the phototrophic and multicellular character of a cyanobacterium, Anabaena.


2004 ◽  
Vol 24 (12) ◽  
pp. 5421-5433 ◽  
Author(s):  
Marco Prisco ◽  
Arianna Maiorana ◽  
Clara Guerzoni ◽  
George Calin ◽  
Bruno Calabretta ◽  
...  

ABSTRACT Pescadillo (PES1) and the upstream binding factor (UBF1) play a role in ribosome biogenesis, which regulates cell size, an important component of cell proliferation. We have investigated the effects of PES1 and UBF1 on the growth and differentiation of cell lines derived from 32D cells, an interleukin-3 (IL-3)-dependent murine myeloid cell line. Parental 32D cells and 32D IGF-IR cells (expressing increased levels of the type 1 insulin-like growth factor I [IGF-I] receptor [IGF-IR]) do not express insulin receptor substrate 1 (IRS-1) or IRS-2. 32D IGF-IR cells differentiate when the cells are shifted from IL-3 to IGF-I. Ectopic expression of IRS-1 inhibits differentiation and transforms 32D IGF-IR cells into a tumor-forming cell line. We found that PES1 and UBF1 increased cell size and/or altered the cell cycle distribution of 32D-derived cells but failed to make them IL-3 independent. PES1 and UBF1 also failed to inhibit the differentiation program initiated by the activation of the IGF-IR, which is blocked by IRS-1. 32D IGF-IR cells expressing PES1 or UBF1 differentiate into granulocytes like their parental cells. In contrast, PES1 and UBF1 can transform mouse embryo fibroblasts that have high levels of endogenous IRS-1 and are not prone to differentiation. Our results provide a model for one of the theories of myeloid leukemia, in which both a stimulus of proliferation and a block of differentiation are required for leukemia development.


2016 ◽  
Author(s):  
Stephen Vadia ◽  
Jessica L. Tse ◽  
Jue D. Wang ◽  
Petra Anne Levin

AbstractNutrients—and by extension biosynthetic capacity—positively impact cell size in organisms throughout the tree of life. In bacteria, cell size is reduced three-fold in response to nutrient starvation or accumulation of the alarmone ppGpp, a global inhibitor of biosynthesis. However, whether biosynthetic capacity as a whole determines cell size or if particular anabolic pathways are more important than others remains an open question. Utilizing a top-down approach, here we identify flux through lipid synthesis as the primary biosynthetic determinant ofEscherichia colicell size. Altering flux through lipid synthesis recapitulated the impact of altering nutrients on cell size and morphology, while defects in other biosynthetic pathways either did not impact size or altered size in a lipid-dependent manner. Together our findings support a model in which lipid availability dictates cell envelope capacity and ppGpp functions as a linchpin linking surface area expansion with cytoplasmic volume to maintain cellular integrity.


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