TY  - JOUR
A1  - Briese, Michael
A1  - Saal, Lena
A1  - Appenzeller, Silke
A1  - Moradi, Mehri
A1  - Baluapuri, Apoorva
A1  - Sendtner, Michael
T1  - Whole transcriptome profiling reveals the RNA content of motor axons
JF  - Nucleic Acids Research
N2  - Most RNAs within polarized cells such as neurons are sorted subcellularly in a coordinated manner. Despite advances in the development of methods for profiling polyadenylated RNAs from small amounts of input RNA, techniques for profiling coding and non-coding RNAs simultaneously are not well established. Here, we optimized a transcriptome profiling method based on double-random priming and applied it to serially diluted total RNA down to 10 pg. Read counts of expressed genes were robustly correlated between replicates, indicating that the method is both reproducible and scalable. Our transcriptome profiling method detected both coding and long non-coding RNAs sized >300 bases. Compared to total RNAseq using a conventional approach our protocol detected 70% more genes due to reduced capture of ribosomal RNAs. We used our method to analyze the RNA composition of compartmentalized motoneurons. The somatodendritic compartment was enriched for transcripts with post-synaptic functions as well as for certain nuclear non-coding RNAs such as 7SK. In axons, transcripts related to translation were enriched including the cytoplasmic non-coding RNA 7SL. Our profiling method can be applied to a wide range of investigations including perturbations of subcellular transcriptomes in neurodegenerative diseases and investigations of microdissected tissue samples such as anatomically defined fiber tracts.
KW  - RNA
KW  - motor axons
Y1  - 2015
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-126800
ER  - 
TY  - JOUR
A1  - Ji, Changhe
A1  - Bader, Jakob
A1  - Ramanathan, Pradhipa
A1  - Hennlein, Luisa
A1  - Meissner, Felix
A1  - Jablonka, Sibylle
A1  - Mann, Matthias
A1  - Fischer, Utz
A1  - Sendtner, Michael
A1  - Briese, Michael
T1  - Interaction of 7SK with the Smn complex modulates snRNP production
JF  - Nature Communications
N2  - Gene expression requires tight coordination of the molecular machineries that mediate transcription and splicing. While the interplay between transcription kinetics and spliceosome fidelity has been investigated before, less is known about mechanisms regulating the assembly of the spliceosomal machinery in response to transcription changes. Here, we report an association of the Smn complex, which mediates spliceosomal snRNP biogenesis, with the 7SK complex involved in transcriptional regulation. We found that Smn interacts with the 7SK core components Larp7 and Mepce and specifically associates with 7SK subcomplexes containing hnRNP R. The association between Smn and 7SK complexes is enhanced upon transcriptional inhibition leading to reduced production of snRNPs. Taken together, our findings reveal a functional association of Smn and 7SK complexes that is governed by global changes in transcription. Thus, in addition to its canonical nuclear role in transcriptional regulation, 7SK has cytosolic functions in fine-tuning spliceosome production according to transcriptional demand.
KW  - Molecular neuroscience
KW  - RNA
KW  - RNA splicing
KW  - Transcription
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259125
VL  - 12
IS  - 1
ER  -