Strikingly, the presence of this frameshifted proteoform was necessary for myocardial contraction, as we showed in PLEKHM2-KO cells that required both the WT and frameshifted proteoforms to recover fully, leading to our model

Furthermore, we were able to show that this frameshifted version of PLEKHM2 generated a hyper-active version of this protein, which did not require activation of its canonical activator

Not only did the frameshift exist, it was highly conserved and encoded for an additional alpha-helix domain

Today our journey ends! We were able to find a highly conserved cellular +1 PRF signal in the gene PLEKHM2/SKIP

Excited to announce the YAK lab's first paper and the discovery of the FIRST human cellular PRF signal to give access to two overlapping open reading frames (science.org/doi/10.1126/...)! Before we dive in, the story actually begins in a Nature from 11 years ago

Thus, any kind of SNARE can be processed universally by this single molecule machine. Check out the paper for more in-depth structural work! (5/5)

Combining in-vivo mass-spec, single molecule FRET, and time resolved cryoEM, we were able to determine that this NSF/Sec18 ring actually splits itself open from the side to accomodate SNAREs, thus completely bypassing any substrate topology issues! (4/5)

The complex that recycles these SNAREs is called NSF/Sec18. However, our mehanistic understanding of how these complex SNARE proteins are pulled through the NSF/Sec18 complex is poor. SNAREs are like a knotted thread that must somehow be threaded by a NSF/Sec18 needle (3/5)

SNARE proteins are the engines for membrane fusion; they cause neurotransmitters to be released in a synpatic cleft all the way to the relase of hormones. After these SNAREs perform their job, they must naturally be recyled and prepared for additional rounds of fusion (2/5)

Sharing some of my PhD work that just came out today.

TLDR; We found a universal mechanism for all the many kinds of SNAREs are processed and recycled by a single molecular machine that is conserved in all eukaryotic life as we know it (1/5)

www.nature.com/articles/s41...

(1/5)

Pretty sure this is a contaminant but do #CryoEM #TeamTomo people know exactly what this is? One of my many 3d classes

What do you make of this? relion.readthedocs.io/en/release-4...

My understanding is that having the falcon4i convert to tiff is equivalent to having relion convert EER to tiff. If we assume that, then we would want to take the inverse of this gain I think?

There are many problems with the current peer review process... but I can't deny that there's a certain feeling when all FOUR (yes, 4) reviewers all unaminiously agree on the quality of your manuscript

2) #RNA elements can dynamically recode the genome: they force the ribosome to change its reading frame dynamically. Viruses employ this routinely (HIV, SARS-CoV2) but this mechanism in humans had been controversial until we found the first human hit in PLKEHM2 (www.biorxiv.org/content/10.1...)

2/3

A proper intro: I'm at Stanford currently and I study (3) things 🧪
1) #cryoEM and #cryoET #teamtomo
of molecular machines: We put out a preprint on how the AAA+ machine Sec18 bypasses topology of all SNARE substrates by using 'side-release' and 'side-loading' (www.biorxiv.org/content/10.1...)
1/3

2) #RNA elements can dynamically recode the genome: they force the ribosome to change its reading frame dynamically. Viruses employ this routinely (HIV, SARS-CoV2) but this mechanism in humans had been controversial until we found the first human hit in PLKEHM2 (www.biorxiv.org/content/10.1...)

2/3

2)RNA elements can dynamically recode the genome: they force the ribosome to change its reading frame dynamically. Viruses employ this routinely (HIV, SARS-CoV2) but this mechanism in humans had been controversial until we found the first human hit in PLKEHM2 (www.biorxiv.org/content/10.1...)

(3/4)

1) We recently put out a preprint about how the AAA+ machine Sec18/NSF (www.biorxiv.org/content/10.1...) are able to load every and any SNARE substrate in the cell by bypassing their N- and C-terminal domains completely by employing a mechanism we've coined 'side-loading' and 'side-release'

(2/4)