Control of gene expression by cell size

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Biology, 2010.This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.Cataloged from student submitted PDF version of thesis.Includes bibliographical references.Polyploidy, increased copy number of whole chromosome sets in the genome, is a common cellular state in evolution, development and disease. Polyploidy enlarges cell size and alters gene expression, producing novel phenotypes and functions. Although many polyploid cell types have been discovered, it is not clear how polyploidy changes physiology. Specifically, whether the enlarged cell size of polyploids causes differential gene regulation has not been investigated. In this thesis, I present the evidence for a size-sensing mechanism that alters gene expression in yeast. My results indicate a causal relationship between cell size and gene expression. Ploidy-associated changes in the transcriptome therefore reflect transcriptional adjustment to a larger cell size. The causal and regulatory connection between cell size and transcription suggests that the physical features of a cell (such as size and shape) are a systematic factor in gene regulation. In addition, cell size homeostasis may have a critical function - maintenance of transcriptional homeostasis.by Chia-Yung Wu.Ph.D

Mlp-dependent anchorage and stabilization of a desumoylating enzyme is required to prevent clonal lethality

Myosin-like proteins 1 and 2 (Mlp1 and Mlp2) form filaments attached to the nucleoplasmic side of the nuclear pore complexes via interaction with the nucleoporin Nup60. Here, we show that Mlps and Nup60, but not several other nucleoporins, are required to localize and stabilize a desumoylating enzyme Ulp1. Moreover, like Mlps, Ulp1 exhibits a unique asymmetric distribution on the nuclear envelope. Consistent with a role in regulating Ulp1, removal of either or both MLPs affects the SUMO conjugate pattern. We also show that deleting MLPs or the localization domains of Ulp1 results in DNA damage sensitivity and clonal lethality, the latter of which is caused by increased levels of 2-micron circle DNA. Epistatic and dosage suppression analyses further demonstrate that Mlps function upstream of Ulp1 in 2-micron circle maintenance and the damage response. Together, our results reveal that Mlps play important roles in regulating Ulp1 and subsequently affect sumoylation stasis, growth, and DNA repair

Regulation of CLC-1 chloride channel biosynthesis by FKBP8 and Hsp90β.

Mutations in human CLC-1 chloride channel are associated with the skeletal muscle disorder myotonia congenita. The disease-causing mutant A531V manifests enhanced proteasomal degradation of CLC-1. We recently found that CLC-1 degradation is mediated by cullin 4 ubiquitin ligase complex. It is currently unclear how quality control and protein degradation systems coordinate with each other to process the biosynthesis of CLC-1. Herein we aim to ascertain the molecular nature of the protein quality control system for CLC-1. We identified three CLC-1-interacting proteins that are well-known heat shock protein 90 (Hsp90)-associated co-chaperones: FK506-binding protein 8 (FKBP8), activator of Hsp90 ATPase homolog 1 (Aha1), and Hsp70/Hsp90 organizing protein (HOP). These co-chaperones promote both the protein level and the functional expression of CLC-1 wild-type and A531V mutant. CLC-1 biosynthesis is also facilitated by the molecular chaperones Hsc70 and Hsp90β. The protein stability of CLC-1 is notably increased by FKBP8 and the Hsp90β inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) that substantially suppresses cullin 4 expression. We further confirmed that cullin 4 may interact with Hsp90β and FKBP8. Our data are consistent with the idea that FKBP8 and Hsp90β play an essential role in the late phase of CLC-1 quality control by dynamically coordinating protein folding and degradation

Simplified ZrTiOx-based RRAM cell structure with rectifying characteristics by integrating Ni/n + -Si diode

A simplified one-diode one-resistor (1D1R) resistive switching memory cell that uses only four layers of TaN/ZrTiO( x )/Ni/n(+)-Si was proposed to suppress sneak current where TaN/ZrTiO( x )/Ni can be regarded as a resistive-switching random access memory (RRAM) device while Ni/n(+)-Si acts as an Schottky diode. This is the first RRAM cell structure that employs metal/semiconductor Schottky diode for current rectifying. The 1D1R cell exhibits bipolar switching behavior with SET/RESET voltage close to 1 V without requiring a forming process. More importantly, the cell shows tight resistance distribution for different states, significantly rectifying characteristics with forward/reverse current ratio higher than 10(3) and a resistance ratio larger than 10(3) between two states. Furthermore, the cell also displays desirable reliability performance in terms of long data retention time of up to 10(4) s and robust endurance of 10(5) cycles. Based on the promising characteristics, the four-layer 1D1R structure holds the great potential for next-generation nonvolatile memory technology

Disordered Fe vacancies and superconductivity in potassium-intercalated iron selenide (K2-xFe4+ySe5)

The parent compound of an unconventional superconductor must contain unusual correlated electronic and magnetic properties of its own. In the high-Tc potassium intercalated FeSe, there has been significant debate regarding what the exact parent compound is. Our studies unambiguously show that the Fe-vacancy ordered K2Fe4Se5 is the magnetic, Mott insulating parent compound of the superconducting state. Non-superconducting K2Fe4Se5 becomes a superconductor after high temperature annealing, and the overall picture indicates that superconductivity in K2-xFe4+ySe5 originates from the Fe-vacancy order to disorder transition. Thus, the long pending question whether magnetic and superconducting state are competing or cooperating for cuprate superconductors may also apply to the Fe-chalcogenide superconductors. It is believed that the iron selenides and related compounds will provide essential information to understand the origin of superconductivity in the iron-based superconductors, and possibly to the superconducting cuprates

Antioxidant activity and growth inhibition of human colon cancer cells by crude and purified fucoidan preparations extracted from Sargassum cristaefolium

AbstractFucose-containing sulfated polysaccharides, also termed “fucoidans”, which are known to possess antioxidant, anticoagulant, anticancer, antiviral, and immunomodulating properties, are normally isolated from brown algae via various extraction techniques. In the present study, two methods (SC1 and SC2) for isolation of fucoidan from Sargassum cristaefolium were compared, with regard to the extraction yields, antioxidant activity, and inhibition of growth of human colon cancer cells exhibited by the respective extracts. SC1 and SC2 differ in the number of extraction steps and concentration of ethanol used, as well as the obtained sulfated polysaccharide extracts, namely, crude fucoidan preparation (CFP) and purified fucoidan preparation (PFP), respectively. Thin layer chromatography, Fourier transform infrared analysis, and measurements of fucose and sulfate contents revealed that the extracts were fucoidan. There was a higher extraction yield for CFP, which contained less fucose and sulfate but more uronic acid, and had weaker antioxidant activity and inhibition of growth in human colon cancer cells. In contrast, there was a lower extraction yield for PFP, which contained more fucose and sulfate but less uronic acid, and had stronger antioxidant activity and inhibition of growth in human colon cancer cells. Thus, since the difference in bioactive activities between CFP and PFP was not remarkable, the high extraction yield of SC1 might be favored as a method in industrial usage for extracting fucoidan

A Study on the Effect of Design Factors of Slim Keyboard's Tactile Feedback

With the rapid development of computer technology, the design of computers and keyboards moves towards a trend of slimness. The change of mobile input devices directly influences users' behavior. Although multi-touch applications allow entering texts through a virtual keyboard, the performance, feedback, and comfortableness of the technology is inferior to traditional keyboard, and while manufacturers launch mobile touch keyboards and projection keyboards, the performance has not been satisfying. Therefore, this study discussed the design factors of slim pressure-sensitive keyboards. The factors were evaluated with an objective (accuracy and speed) and a subjective evaluation (operability, recognition, feedback, and difficulty) depending on the shape (circle, rectangle, and L-shaped), thickness (flat, 3mm, and 6mm), and force (35±10g, 60±10g, and 85±10g) of the keyboard. Moreover, MANOVA and Taguchi methods (regarding signal-to-noise ratios) were conducted to find the optimal level of each design factor. The research participants, by their typing speed (30 words/ minute), were divided in two groups. Considering the multitude of variables and levels, the experiments were implemented using the fractional factorial design. A representative model of the research samples were established for input task testing. The findings of this study showed that participants with low typing speed primarily relied on vision to recognize the keys, and those with high typing speed relied on tactile feedback that was affected by the thickness and force of the keys. In the objective and subjective evaluation, a combination of keyboard design factors that might result in higher performance and satisfaction was identified (L-shaped, 3mm, and 60±10g) as the optimal combination. The learning curve was analyzed to make a comparison with a traditional standard keyboard to investigate the influence of user experience on keyboard operation. The research results indicated the optimal combination provided input performance to inferior to a standard keyboard. The results could serve as a reference for the development of related products in industry and for applying comprehensively to touch devices and input interfaces which are interacted with people

A Study on the Effect of Design Factors of Slim Keyboard's Tactile Feedback

With the rapid development of computer technology, the design of computers and keyboards moves towards a trend of slimness. The change of mobile input devices directly influences users' behavior. Although multi-touch applications allow entering texts through a virtual keyboard, the performance, feedback, and comfortableness of the technology is inferior to traditional keyboard, and while manufacturers launch mobile touch keyboards and projection keyboards, the performance has not been satisfying. Therefore, this study discussed the design factors of slim pressure-sensitive keyboards. The factors were evaluated with an objective (accuracy and speed) and a subjective evaluation (operability, recognition, feedback, and difficulty) depending on the shape (circle, rectangle, and L-shaped), thickness (flat, 3mm, and 6mm), and force (35±10g, 60±10g, and 85±10g) of the keyboard. Moreover, MANOVA and Taguchi methods (regarding signal-to-noise ratios) were conducted to find the optimal level of each design factor. The research participants, by their typing speed (30 words/ minute), were divided in two groups. Considering the multitude of variables and levels, the experiments were implemented using the fractional factorial design. A representative model of the research samples were established for input task testing. The findings of this study showed that participants with low typing speed primarily relied on vision to recognize the keys, and those with high typing speed relied on tactile feedback that was affected by the thickness and force of the keys. In the objective and subjective evaluation, a combination of keyboard design factors that might result in higher performance and satisfaction was identified (L-shaped, 3mm, and 60±10g) as the optimal combination. The learning curve was analyzed to make a comparison with a traditional standard keyboard to investigate the influence of user experience on keyboard operation. The research results indicated the optimal combination provided input performance to inferior to a standard keyboard. The results could serve as a reference for the development of related products in industry and for applying comprehensively to touch devices and input interfaces which are interacted with people