SASDDY4: Protein sex-lethal mutant with 10GS-linker (Protein sex-lethal mu...

Basic information

• Entry: Database: SASBDB / ID: SASDDY4

Sample

Protein sex-lethal mutant with 10GS-linker

• Protein sex-lethal mutant (protein), Sxl10GS, Drosophila melanogaster

Function / homology

Function:

sex determination, primary response to X:A ratio / germarium-derived cystoblast division / epithelium regeneration / female sex determination / somatic sex determination / female germ-line sex determination / oocyte differentiation / imaginal disc growth / negative regulation of RNA export from nucleus / sex-chromosome dosage compensation / regulation of stem cell division / sex determination / poly-pyrimidine tract binding / sex differentiation / alternative mRNA splicing, via spliceosome / negative regulation of receptor signaling pathway via JAK-STAT / poly(A) binding / pre-mRNA binding / positive regulation of smoothened signaling pathway / regulation of mRNA splicing, via spliceosome / reciprocal meiotic recombination / poly(U) RNA binding / oogenesis / regulation of alternative mRNA splicing, via spliceosome / negative regulation of mRNA splicing, via spliceosome / negative regulation of translational initiation / mRNA regulatory element binding translation repressor activity / positive regulation of RNA splicing / mRNA 3'-UTR binding / mRNA 5'-UTR binding / negative regulation of translation / protein stabilization / ribonucleoprotein complex / mRNA binding / protein-containing complex / RNA binding / nucleus / cytosol / cytoplasm

Domain/homology:

Sex-lethal splicing factor / Paraneoplastic encephalomyelitis antigen / RNA recognition motif / RNA recognition motif / Eukaryotic RNA Recognition Motif (RRM) profile. / RNA recognition motif domain / RNA-binding domain superfamily / Nucleotide-binding alpha-beta plait domain superfamily

Component:

Protein sex-lethal

• Biological species: Drosophila melanogaster (fruit fly)
• Citation: Journal: ACS Comb Sci / Year: 2018; Title: A General Small-Angle X-ray Scattering-Based Screening Protocol Validated for Protein-RNA Interactions.; Authors: Po-Chia Chen / Pawel Masiewicz / Vladimir Rybin / Dmitri Svergun / Janosch Hennig / ; Abstract: We present a screening protocol utilizing small-angle X-ray scattering (SAXS) to obtain structural information on biomolecular interactions independent of prior knowledge, so as to complement affinity-based screening and provide leads for further exploration. This protocol categorizes ligand titrations by computing pairwise agreement between curves, and separately estimates affinities by quantifying complex formation as a departure from the linear sum properties of solution SAXS. The protocol is validated by sparse sequence search around the native poly uridine RNA motifs of the two-RRM domain Sex-lethal protein (Sxl). The screening of 35 RNA motifs between 4 to 10 nucleotides reveals a strong variation of resulting complexes, revealed to be preference-switching between 1:1 and 2:2 binding stoichiometries upon addition of structural modeling. Validation of select sequences in isothermal calorimetry and NMR titration retrieves domain-specific roles and function of a guanine anchor. These findings reinforce the suitability of SAXS as a complement in lead identification.

Contact author

• Po-chia Chen (EMBL, European Molecular Biology Laboratory (EMBL) - Heidelberg, Heidelberg, Germany)

Models

• Model #1866: ; Type: mix / Software: (2.1) / Radius of dummy atoms: 1.90 A / Chi-square value: 0.679 / P-value: 0.675072; Search similar-shape structures of this assembly by Omokage search (details)
• Model #1867: ; Type: mix / Software: (2.1) / Radius of dummy atoms: 1.90 A / Chi-square value: 0.679 / P-value: 0.675072; Search similar-shape structures of this assembly by Omokage search (details)
• Model #1868: ; Type: mix / Software: (2.1) / Radius of dummy atoms: 1.90 A / Chi-square value: 0.679 / P-value: 0.675072; Search similar-shape structures of this assembly by Omokage search (details)
• Model #1869: ; Type: mix / Software: (2.1) / Radius of dummy atoms: 1.90 A / Chi-square value: 0.679 / P-value: 0.675072; Search similar-shape structures of this assembly by Omokage search (details)
• Model #1870: ; Type: mix / Software: (2.1) / Radius of dummy atoms: 1.90 A / Chi-square value: 0.679 / P-value: 0.675072; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Protein sex-lethal mutant with 10GS-linker / Specimen concentration: 1 mg/ml
• Buffer: Name: 10 mM KP, 50 mM NaCl, 10 mM DTT / pH: 6

Entity #1006

Name: Sxl10GS / Type: protein / Description: Protein sex-lethal mutant / Formula weight: 20.432 / Num. of mol.: 1 / Source: Drosophila melanogaster / References: UniProt: P19339
Sequence:

GAMASNTNLI VNYLPQDMTD RELYALFRAI GPINTCRIMR DYKTGYSFGY AFVDFTSEMD SQRAIKVLNG ITVRNKRLKV SYARPGGGSG SGGGGSGESI KDTNLYVTNL PRTITDDQLD TIFGKYGSIV QKNILRDKLT GRPRGVAFVR YNKREEAQEA ISALNNVIPE GGSQPLSVRL AEEHGK

Experimental information

• Beam: Instrument name: ESRF BM29 / City: Grenoble / 国: France / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.09919 Å / Dist. spec. to detc.: 2.867 mm
• Detector: Name: Pilatus 1M

Scan

Title: Protein sex-lethal mutant with 10GS-linker / Measurement date: Feb 2, 2017 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 1 sec. / Number of frames: 10 / Unit: 1/nm /

	Min	Max
Q	0.0348	4.9417

Distance distribution function P(R)

Sofotware P(R): GNOM 4.5a / Number of points: 297 /

	Min	Max
Q	0.2114	2.997
P(R) point	1	297
R	0	7.398

Result

Type of curve: single_conc /

	Experimental	Porod
MW	16 kDa	15.85 kDa
Volume	-	26.95 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	22.77	0.05	22.66	0.02
Radius of gyration, Rg	2.18 nm	0.009	2.12 nm	0.05

	Min	Max
D	-	7.4
Guinier point	12	121