Structural basis of the recognition of adeno-associated virus by the neurological system-related receptor carbonic anhydrase IV.

Carbonic anhydrase IV (Car4) is a newly identified receptor that allows adeno-associated virus (AAV) 9P31 to cross the blood-brain barrier and achieve efficient infection in the central nervous system (CNS) in mouse models. However, the molecular mechanism by which engineered AAV capsids with 7-mer insertion in the variable region (VR) VIII recognize these novel cellular receptors is unknown. Here we report the cryo-EM structures of AAV9P31 and its complex with Mus musculus Car4 at atomic resolution by utilizing the block-based reconstruction (BBR) method. The structures demonstrated that Car4 binds to the protrusions at 3-fold axes of the capsid. The inserted 7-mer extends into a hydrophobic region near the catalytic center of Car4 to form stable interactions. Mutagenesis studies also identified the key residues in Car4 responsible for the AAV9P31 interaction. These findings provide new insights into the novel receptor recognition mechanism of AAV generated by directed evolution and highlight the application of the BBR method to studying the virus-receptor molecular mechanism.

A mechanism for SARS-CoV-2 RNA capping and its inhibition by nucleotide analog inhibitors.

Decoration of cap on viral RNA plays essential roles in SARS-CoV-2 proliferation. Here, we report a mechanism for SARS-CoV-2 RNA capping and document structural details at atomic resolution. The NiRAN domain in polymerase catalyzes the covalent link of RNA 5' end to the first residue of nsp9 (termed as RNAylation), thus being an intermediate to form cap core (GpppA) with GTP catalyzed again by NiRAN. We also reveal that triphosphorylated nucleotide analog inhibitors can be bonded to nsp9 and fit into a previously unknown "Nuc-pocket" in NiRAN, thus inhibiting nsp9 RNAylation and formation of GpppA. S-loop (residues 50-KTN-52) in NiRAN presents a remarkable conformational shift observed in RTC bound with sofosbuvir monophosphate, reasoning an "induce-and-lock" mechanism to design inhibitors. These findings not only improve the understanding of SARS-CoV-2 RNA capping and the mode of action of NAIs but also provide a strategy to design antiviral drugs.

Frequency-modulated continuous-wave laser ranging using low-duty-cycle signals for the applications of real-time super-resolved ranging.

A frequency-modulated continuous-wave laser ranging method using low-duty-cycle linear-frequency-modulated (LFM) signals is proposed. A spectrum consisting of a dense Kronecker comb is obtained so that the frequency of the beat signal can be measured with finer resolution. Since the dense comb is provided, super-resolved laser ranging can be achieved using a single-parametric frequency estimation method. Therefore, the run times of the estimation are reduced which promises real-time applications. A proof-of-concept experiment is carried out, in which an LFM signal with a bandwidth of 5 GHz and a duration of 1 µs is used. The duty-cycle of the LFM signal is 10%. The time delay of a scanning variable optical delay line is obtained in real time from the frequency of the highest comb tooth, of which the measurement resolution is 20 ps. Moreover, a single-parametric nonlinear least squares method is used to fit the envelope so that the time delay can be estimated with super-resolution. The standard deviation of the estimation displacements is 2.3 ps, which is 87 times finer than the bandwidth-limited resolution (200 ps). Therefore, the variation of the time delay can be precisely monitored. The proposed method may be used to achieve real-time high-resolution laser ranging with low-speed electronic devices.