Dietary sources of thiamine (vitamin B 1 ) and thiamine-degrading enzymes (thiaminases) are thought to be primary factors in the development of thiamine deficiency among Great Lakes salmonines. We surveyed major forage fish species in Lake Michigan for their content of thiamine, thiamine vitamers, and thiaminase activity. Concentrations of total thiamine were similar (P Յ 0.05) among most forage fishes (alewife Alosa pseudoharengus, bloater Coregonus hoyi, spottail shiner Notropis hudsonius, deepwater sculpin Myoxocephalus thompsonii, yellow perch Perca flavescens, ninespine stickleback Pungitius pungitius, and round goby Neogobius melanostomus) and slightly lower in rainbow smelt Osmerus mordax. Concentrations of total thiamine were all above the dietary requirements of coldwater fishes, suggesting the thiamine content of forage fish is not the critical factor in the development of thiamine deficiency in Lake Michigan salmonines. Thiamine pyrophosphate was the predominant form of thiamine in most species of forage fish, followed by free thiamine and thiamine monophosphate. Total thiamine was slightly greater in summer collections of alewife and rainbow smelt than in spring and fall collections, but the same was not true for bloater. Thiaminase activity varied among species and was greatest in gizzard shad Dorosoma cepedianum, spottail shiner, alewife, and rainbow smelt. Thiaminase activity in alewife varied among collection locations, season (greatest in spring), and size of the fish. Size and condition factors were positively correlated with both total thiamine and thiaminase activity in alewife. Thus, thiamine and thiaminase activity in forage fishes collected in Lake Michigan varied among species, seasons, year caught, and size (or condition). Therefore, multiple factors must be considered in the development of predictive models for the onset of thiamine deficiency in Great Lakes salmonines. Most importantly, thiaminase activity was great in alewives and rainbow smelt, suggesting that these prey fish are key causative factors of the thiamine deficiency in Great Lakes salmonines.

Microrchidia 3 (MORC3) is a human protein linked to autoimmune disorders, Down syndrome, and cancer. It is a member of a newly identified family of human ATPases with an uncharacterized mechanism of action. Here, we elucidate the molecular basis for inhibition and activation of MORC3. The crystal structure of the MORC3 region encompassing the ATPase and CW domains in complex with a nonhydrolyzable ATP analog demonstrates that the two domains are directly coupled. The extensive ATPase:CW interface stabilizes the protein fold but inhibits the catalytic activity of MORC3. Enzymatic, NMR, mutational, and biochemical analyses show that in the autoinhibited, off state, the CW domain sterically impedes binding of the ATPase domain to DNA, which in turn is required for the catalytic activity. MORC3 autoinhibition is released by disrupting the intramolecular ATPase:CW coupling through the competitive interaction of CW with histone H3 tail or by mutating the interfacial residues. Binding of CW to H3 leads to a marked rearrangement in the ATPase–CW cassette, which frees the DNA-binding site in active MORC3 (on state). We show that ATP-induced dimerization of the ATPase domain is strictly required for the catalytic activity and that the dimeric form of ATPase–CW might cooperatively bind to dsDNA. Together, our findings uncovered a mechanism underlying the fine-tuned regulation of the catalytic domain of MORC3 by the epigenetic reader, CW.

No abstract

Phase separation can produce local structures with specific functionality in the cell, and in the nucleus, this can lead to chromatin reorganization. Microrchidia 3 (MORC3) is a human ATPase that has been implicated in autoimmune disorders and cancer. Here, we show that MORC3 forms phase-separated condensates with liquid-like properties in the cell nucleus. Fluorescence live-cell imaging reveals that the MORC3 condensates are heterogeneous and undergo dynamic morphological changes during the cell cycle. The ATPase activity of MORC3 drives its phase separation in vitro and requires DNA binding and releasing the MORC3 CW domain-dependent autoinhibition through association with histone H3. Our findings suggest a mechanism by which the ATPase function of MORC3 mediates MORC3 nuclear compartmentalization.

Nucleosomal DNA sequences generally follow a well-known pattern with ∼10-bp periodic WW (where W is A or T) dinucleotides that oscillate in phase with each other and out of phase with SS (where S is G or C) dinucleotides. However, nucleosomes with other DNA patterns have not been systematically analyzed. Here, we focus on an opposite pattern, namely anti-WW/SS pattern, in which WW dinucleotides preferentially occur at DNA sites that bend into major grooves and SS (where S is G or C) dinucleotides are often found at sites that bend into minor grooves. Nucleosomes with the anti-WW/SS pattern are widespread and exhibit a species- and context-specific distribution in eukaryotic genomes. Unlike non-mammals (yeast, nematode and fly), there is a positive correlation between the enrichment of anti-WW/SS nucleosomes and RNA Pol II transcriptional levels in mammals (mouse and human). Interestingly, such enrichment is not due to underlying DNA sequence. In addition, chromatin remodeling complexes have an impact on the abundance but not on the distribution of anti-WW/SS nucleosomes in yeast. Our data reveal distinct roles of cis- and trans-acting factors in the rotational positioning of nucleosomes between non-mammals and mammals. Implications of the anti-WW/SS sequence pattern for RNA Pol II transcription are discussed.