A spatiotemporal map of co-receptor signaling networks underlying B cell activation.

The B cell receptor (BCR) signals together with a multi-component co-receptor complex to initiate B cell activation in response to antigen binding. Here, we take advantage of peroxidase-catalyzed proximity labeling combined with quantitative mass spectrometry to track co-receptor signaling dynamics in Raji cells from 10 s to 2 h after BCR stimulation. This approach enables tracking of 2,814 proximity-labeled proteins and 1,394 phosphosites and provides an unbiased and quantitative molecular map of proteins recruited to the vicinity of CD19, the signaling subunit of the co-receptor complex. We detail the recruitment kinetics of signaling effectors to CD19 and identify previously uncharacterized mediators of B cell activation. We show that the glutamate transporter SLC1A1 is responsible for mediating rapid metabolic reprogramming and for maintaining redox homeostasis during B cell activation. This study provides a comprehensive map of BCR signaling and a rich resource for uncovering the complex signaling networks that regulate activation.

Catching proteins for degradation.

Aubiquitin-independent pathway targets nuclear proteins to the proteasome.

A Spatiotemporal Map of Co-Receptor Signaling Networks Underlying B Cell Activation.

The B cell receptor (BCR) signals together with a multi-component co-receptor complex to initiate B cell activation in response to antigen binding. This process underlies nearly every aspect of proper B cell function. Here, we take advantage of peroxidase-catalyzed proximity labeling combined with quantitative mass spectrometry to track B cell co-receptor signaling dynamics from 10 seconds to 2 hours after BCR stimulation. This approach enables tracking of 2,814 proximity-labeled proteins and 1,394 quantified phosphosites and provides an unbiased and quantitative molecular map of proteins recruited to the vicinity of CD19, the key signaling subunit of the co-receptor complex. We detail the recruitment kinetics of essential signaling effectors to CD19 following activation, and then identify new mediators of B cell activation. In particular, we show that the glutamate transporter SLC1A1 is responsible for mediating rapid metabolic reprogramming immediately downstream of BCR stimulation and for maintaining redox homeostasis during B cell activation. This study provides a comprehensive map of the BCR signaling pathway and a rich resource for uncovering the complex signaling networks that regulate B cell activation.

Transcriptional control of mycobacterial DNA damage response by sigma adaptation.

Transcriptional activator PafBC is the key regulator of the mycobacterial DNA damage response and controls around 150 genes, including genes involved in the canonical SOS response, through an unknown molecular mechanism. Using a combination of biochemistry and cryo­electron microscopy, we demonstrate that PafBC in the presence of single-stranded DNA activates transcription by reprogramming the canonical −10 and −35 promoter specificity of RNA polymerase associated with the housekeeping sigma subunit. We determine the structure of this transcription initiation complex, revealing a unique mode of promoter recognition, which we term "sigma adaptation." PafBC inserts between DNA and sigma factor to mediate recognition of hybrid promoters lacking the −35 but featuring the canonical −10 and a PafBC-specific −26 element. Sigma adaptation may constitute a more general mechanism of transcriptional control in mycobacteria.

A yellow chlorophyll catabolite in leaves of Urtica dioica L.: An overlooked phytochemical that contributes to health benefits of stinging nettle.

Stinging nettle is appreciated for its antioxidant and anti-inflammatory properties, which renders the plant a popular ingredient in a healthy diet in form of salads or smoothies. The most common use, presumably, is of dried leaves as ingredient in tea mixtures. The plant's health benefits are attributed primarily to phenolic phytochemicals. Here we describe the characterization and quantification of a phylloxanthobilin (PxB), a yellow chlorophyll catabolite, in nettle tea. Despite their abundance in the plant kingdom, chlorophyll catabolites have been overlooked as phytochemicals and as part of human nutrition. Our investigations of tea reveal that one cup of nettle tea contains about 50 µg of PxB with large variations depending on the supplier. When investigating the bioactivities of PxB, our observations show that PxB has antioxidative and anti-inflammatory activities comparable to known bioactive small molecules found in nettle, indicating the phylloxanthobilin to be an overlooked ingredient of nettle tea.