#nadia plays nms

[iteration-orpheus]

A wonderful story about my time playing NMS is the time in which I realized everyone was they/them. At first I played the game in Spanish, and the use of a neutral pronoun like 'elle' is very uncommon, unfortunately, so everyone used he/him. But since I was stupid, when meeting Nada and Polo, I assumed that they were a straight couple (since Nada sounds similar to Nadia, a female name), and insulted the fact that the only 'female' character was there to avoid gay. Then I realized Nada used he/him (in the esp version) and got very happy for gay. Then I switched to playing in English and saw that everyone used they/them and got even happier (I screeched and my sibling got scared) . Curse whoever misgendered my aliens in the Spanish version, but thanks to that I got the suprise of my life

[softgrungeprophet]

did not know that no man’s sky had like, fire in it, but apparently it does

[naivelocus]

Cryo-EM Reveals How Human Cytoplasmic Dynein Is Auto-inhibited and Activated

Highlights

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Cryo-EM shows human cytoplasmic dynein-1 in its auto-inhibited, phi-particle form

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Phi-particle disruption in vitro and in cells reveals its role in dynein regulation

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There is a transition from phi-particle to open-dynein: both forms are inhibited

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Dynactin binds open-dynein and aligns its motors to activate processive movement

Summary

Cytoplasmic dynein-1 binds dynactin and cargo adaptor proteins to form a transport machine capable of long-distance processive movement along microtubules. However, it is unclear why dynein-1 moves poorly on its own or how it is activated by dynactin. Here, we present a cryoelectron microscopy structure of the complete 1.4-megadalton human dynein-1 complex in an inhibited state known as the phi-particle. We reveal the 3D structure of the cargo binding dynein tail and show how self-dimerization of the motor domains locks them in a conformation with low microtubule affinity. Disrupting motor dimerization with structure-based mutagenesis drives dynein-1 into an open form with higher affinity for both microtubules and dynactin. We find the open form is also inhibited for movement and that dynactin relieves this by reorienting the motor domains to interact correctly with microtubules. Our model explains how dynactin binding to the dynein-1 tail directly stimulates its motor activity.

Keywords

motor;

dynein;

dynactin;

cryo-EM;

microtubule;

phi-particle;

auto-inhibition;

activation

Introduction

Cytoplasmic dynein-1 (dynein) associates with dynactin to form an efficient microtubule motor that transports cargo to the minus end of microtubules and organizes the internal components of eukaryotic cells. Disruption of this 2.4-megadalton machine disperses the Golgi network (Burkhardt et al., 1997), blocks transport between organelles (Presley et al., 1997), and leaves viruses stuck at the cell periphery (Döhner et al., 2002). In addition, dynein and dynactin are required during cell division for spindle formation and correct chromosome alignment (Echeverri et al., 1996). Dynein must therefore be carefully regulated to ensure the correct timing and location of motor activation.

In cells, most dynein is diffuse in the cytoplasm, with only a small fraction on microtubules (Splinter et al., 2012). This prevents dynein from inappropriately saturating microtubules or traveling unnecessarily and ensures there is a pool of dynein ready to transport cargos when required. The switch of dynein and dynactin from diffuse to actively transporting cargo is controlled at many levels. It can be driven both in vitro (Mallik et al., 2005) and in vivo (Rai et al., 2013) by clustering motors and influenced by targeting dynein/dynactin to specific post-translationally modified microtubules (McKenney et al., 2016 ;  Nirschl et al., 2016) or the microtubule plus ends (Duellberg et al., 2014 ;  Moughamian et al., 2013). The switch is also controlled at the level of the dynein/dynactin machinery itself. Whereas isolated human dynein is weakly processive in vitro (Trokter et al., 2012), it can be activated to move over long distances (>500 nm) by binding to dynactin and a cargo-specific adaptor protein such as BICD2 (McKenney et al., 2014 ;  Schlager et al., 2014) or Hook3 (McKenney et al., 2014; Olenick et al., 2016 ;  Schroeder and Vale, 2016). This binding stimulates processive movement by increasing the run length, velocity (McKenney et al., 2014 ;  Schlager et al., 2014), and force output (Belyy et al., 2016) of individual motors.

Dynein consists of two motor domains that are responsible for ATP hydrolysis and force production and a tail region that holds them together. It is unclear why dynein is only weakly processive on its own and how it is activated by dynactin and cargo adaptors. There is some evidence that dynein processivity is directly inhibited by the C-terminal ∼300 amino acids of the motor domain (Nicholas et al., 2015). Another theory is that inhibition is due to the tail region folding back to contact the motor domains until cargo binds (Belyy et al., 2016 ;  Markus et al., 2009). A similar inhibition mechanism is used by cytoskeletal motors in the kinesin (Kaan et al., 2011) and myosin families (Hammer and Sellers, 2011). Alternatively, it has been proposed that dynein is auto-inhibited by self-dimerization of its motor domains (Torisawa et al., 2014). This form of dynein is referred to as the phi-particle because of its resemblance to the Greek letter phi (φ) (Amos, 1989). Activation was suggested to result from a shift in the equilibrium of dynein conformations toward an open form in which the motor domains are separated. In support of this, forced separation of isolated dynein motor domains can increase motor activity (Torisawa et al., 2014). However, these studies were performed on artificially dimerized dynein motors lacking the tail region. It is therefore not clear whether the tail contributes to inhibition or what role the phi-particle plays in the context of the whole dynein complex.

In this study, we set out to determine whether the phi-particle contributes to dynein auto-inhibition and how dynein is activated by dynactin. We use cryoelectron microscopy (cryo-EM) to determine the structure of the phi-particle. We show how the motor domains self-dimerize and are locked in a conformation with weak affinity for microtubules. Disrupting the motor dimer by structure-based mutagenesis drives dynein into an open form with increased affinity for microtubules and dynactin. Surprisingly, we discover that the open form of dynein is also inhibited. We use a combination of 2D analysis of EM images and a 3D cryo-EM structure of the whole dynein/dynactin machinery to explain how dynactin overcomes this inhibition and directly reorients the motor domains to make dynein processive. Finally, we show that disrupting the phi-particle in cells causes mis-localization and mitotic defects, supporting a physiological role for the phi-particle in dynein regulation.

Author : Kai Zhang, Helen E. Foster, Arnaud Rondelet, Samuel E. Lacey, Nadia Bahi-Buisson, Alexander W. Bird, Andrew P. Carter

— Cell

[softgrungeprophet]

Very cool place to be

[softgrungeprophet]

me: oh i wonder what a “hexagonal planet” is, it looks round to me

me a few seconds later: ah

[softgrungeprophet]

built a car, found a lake, found an abyssal horror???? and also some frogtopuses.

[softgrungeprophet]

info on my flying brick of a ship, which the game has designated an Isshi AQ1, presumably no relation to the Isshi ML5 exotic lol

As you can see, I have renamed it the Kingdom Come, after the ship(s) in Friends at the Table, cause it reminds me of jdq’s description from c/W of “two ships stacked on top of each other”

idk how coordinates or galaxies or anything work but I originally traded an NPC alien for it at Mehullk station minor in the Mehullk galaxy (a vy’keen system, i think), region: Tabiko Conflux. PLUS I just saw another one today (pictured in the bottommost screenshot)

The signal booster I built on a nearby planet in the system said “049C:007C:01E0:0134″

I just thought it was interesting cause it’s a very... unique... looking ship and I can’t find any info about it anywhere! I don’t even know what type of starship it is. I think it’s a hauler based on the model... 22 inventory slots + 5 tech slots...

[softgrungeprophet]

followed a distress beacon, found a damaged ship, accidentally parked inside of it, repaired part of it, went to the space station to sell some shit, talked to an alien and traded in my mostly functional but full of garbage fixer-upper for a clean ship without a bunch of junk cluttering the inventory slots

boy does the new one look like two ships stacked on top of each other XD but it’s a B-class instead of C and has like twice the inventory space... so... i’m happy with that

[softgrungeprophet]

something ominous about this place.

[softgrungeprophet]

hmmmmmmmmmmm...

[softgrungeprophet]

strawberry lemonade

[softgrungeprophet]

bro, this shit’s so pretty

[softgrungeprophet]

no man’s sky just reminding me that i haven’t finished soma >>;;

[softgrungeprophet]

for the past few weeks i haven’t been able to get my saved games in NMS to load at all, just a black screen with only the map markers visible, and some UI sometimes...

did some fiddling....

still can’t load my games but now it has the visor hud up???

i’m really bummed cause even if it was “only” a 16 hour save i really liked this game and now it just won’t run at all. i tried changing the window mode, tried compatibility mode, deleted cache stuff, tried to make a new save, and none of it will work. :/

idk what to do about it... i just can’t play this game now i guess...

[softgrungeprophet]

i’ve been messing with some stuff so i changed the suit colors i was using in NMS to see how it looks.... love me some turquoise.

[softgrungeprophet]

sodium-rich planet