Mutations in SPARTAN are associated with early onset hepatocellular carcinoma and progeroid features. A regulatory function of Spartan has been implicated in DNA damage tolerance pathways such as translesion synthesis, but the exact function of the protein remained unclear. Here, we reveal the role of human Spartan in facilitating replication of DNA–protein crosslink-containing DNA. We found that purified Spartan has a DNA-dependent protease activity degrading certain proteins bound to DNA. In concert, Spartan is required for direct DPC removal in vivo; we also show that the protease Spartan facilitates repair of formaldehyde-induced DNA–protein crosslinks in later phases of replication using the bromodeoxyuridin (BrdU) comet assay. Moreover, DNA fibre assay indicates that formaldehyde-induced replication stress dramatically decreases the speed of replication fork movement in Spartan-deficient cells, which accumulate in the G2/M cell cycle phase. Finally, epistasis analysis mapped these Spartan functions to the RAD6-RAD18 DNA damage tolerance pathway. Our results reveal that Spartan facilitates replication of DNA–protein crosslink-containing DNA enzymatically, as a protease, which may explain its role in preventing carcinogenesis and aging.

To date, several algorithms for the retrieval of cyanobacterial phycocyanin (PC) from ocean colour sensors have been presented for inland waters, all of which claim to be robust models. To address this, we conducted a comprehensive comparison to identify the optimal algorithm for retrieval of PC concentrations in the highly optically complex waters of Lake Balaton (Hungary). MEdium Resolution Imaging Spectrometer (MERIS) top-of-atmosphere radiances were first atmospherically corrected using the Self-Contained Atmospheric Parameters Estimation for MERIS data v.B2 (SCAPE-M_B2). Overall, the Simis05 semi-analytical algorithm outperformed more complex inversion algorithms, providing accurate estimates of PC up to ±7 days from the time of satellite overpass during summer cyanobacteria blooms (RMSElog < 0.33). Same-day retrieval of PC also showed good agreement with cyanobacteria biomass (R2 > 0.66, p < 0.001). In-depth analysis of the Simis05 algorithm using in situ measurements of inherent optical properties (IOPs) revealed that the Simis05 model overestimated the phytoplankton absorption coefficient [aph(λ)] by a factor of ~2. However, these errors were compensated for by underestimation of the mass-specific chlorophyll absorption coefficient [a*chla(λ)]. This study reinforces the need for further validation of algorithms over a range of optical water types in the context of the recently launched Ocean Land Colour Instrument (OLCI) onboard Sentinel-3.

We have developed a simple method called I-Block assay, which can detect sequence-specific binding of proteins to DNA in Escherichia coli. The method works by detecting competition between the protein of interest and RNA polymerase for binding to overlapping target sites in a plasmid-borne lacI promoter variant. The assay utilizes two plasmids and an E. coli host strain, from which the gene of the Lac repressor (lacI) has been deleted. One of the plasmids carries the lacI gene with a unique NheI restriction site created in the lacI promoter. The potential recognition sequences of the tested protein are inserted into the NheI site. Introduction of the plasmids into the E. coliΔlacI host represses the constitutive β-galactosidase synthesis of the host bacterium. If the studied protein expressed from a compatible plasmid binds to its target site in the lacI promoter, it will interfere with lacI transcription and lead to increased β-galactosidase activity. The method was tested with two zinc finger proteins, with the lambda phage cI857 repressor, and with CRISPR-dCas9 targeted to the lacI promoter. The I-Block assay was shown to work with standard liquid cultures, with cultures grown in microplate and with colonies on X-gal indicator plates.

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