Recombinant Drosophila melanogaster DNA-directed RNA polymerase II subunit RPB9 (RpII15)

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Cat. No.

BT49278

Source

Yeast

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Purity

>85% (SDS-PAGE)

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Cat. No.

BT49278

Source

E.coli

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Purity

>85% (SDS-PAGE)

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Cat. No.

BT49278

Source

E.coli

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Purity

>85% (SDS-PAGE)

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Cat. No.

BT49278

Source

Baculovirus

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Purity

>85% (SDS-PAGE)

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Cat. No.

BT49278

Source

Mammalian cell

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Purity

>85% (SDS-PAGE)

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Description

Functional Roles in Transcription

RpII15 is indispensable for accurate transcription initiation and elongation:

Start site selection: Loss of RpII15 in Drosophila shifts transcription initiation upstream, mirroring defects observed in yeast ΔRPB9 strains .
Fidelity control: Prevents transcriptional errors by stabilizing the DNA-RNA hybrid at the active site .
Promoter-specific regulation: Binds Gcn4-responsive elements to activate starvation-induced genes like ATG1 (autophagy-related) .

Key Finding: Recombinant RpII15 restores wild-type transcription initiation patterns in vitro when added to RPB9-deficient Pol II .

Recombinant Production and Applications

Recombinant RpII15 is typically expressed in E. coli with affinity tags (e.g., GST or His6) for purification. Functional assays include:

In vitro transcription rescue: 70 ng of recombinant RpII15 corrects aberrant start sites in RPB9-deficient extracts .
DNA-binding assays: EMSA confirms indirect promoter binding via transcription factors like Gcn4 .

Table 2: Biochemical Properties of Recombinant RpII15

Property	Value	Method
Molecular weight	14 kDa	SDS-PAGE
Zinc content	2 atoms/molecule	ICP-MS
Thermostability	Denatures at 65°C	CD spectroscopy

Phenotypic Consequences of RpII15 Deficiency

Lethality: Drosophila RpII15-null mutants are inviable, contrasting with yeast viability under standard conditions .
Temperature sensitivity: Growth defects at extreme temperatures due to impaired Pol II processivity .
Autophagy defects: Reduced ATG1 transcription under nutrient stress, conserved in mammalian cells .

Evolutionary and Mechanistic Insights

Conservation: RpII15 homologs in archaea and eukaryotes share structural roles in Pol II assembly .
Allosteric regulation: RpII15 stabilizes the Pol II clamp domain in a closed conformation, preventing premature DNA entry .
CTD interaction: Facilitates recruitment of phosphatases to recycle Pol II during termination .

Open Questions and Future Directions

Role in elongation: While RpII15 is critical for initiation, its impact on Pol II elongation rates in Drosophila remains uncharacterized .
Disease relevance: Human RPB9 mutations correlate with cancers; analogous studies in Drosophila could model transcriptional dysregulation.

Product Specs

Form

Lyophilized powder. We will ship the in-stock format preferentially. If you have specific format requirements, please note them when ordering, and we will fulfill your request.

Lead Time

Delivery times vary depending on the purchase method and location. Consult your local distributor for specific delivery times. All proteins are shipped with standard blue ice packs. Contact us in advance for dry ice shipping, which incurs extra fees.

Notes

Avoid repeated freezing and thawing. Working aliquots can be stored at 4°C for up to one week.

Reconstitution

Briefly centrifuge the vial before opening to collect contents at the bottom. Reconstitute the protein in sterile deionized water to a concentration of 0.1-1.0 mg/mL. Add 5-50% glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final glycerol concentration is 50%.

Shelf Life

Shelf life depends on several factors, including storage conditions, buffer composition, storage temperature, and protein stability. Generally, the liquid form has a shelf life of 6 months at -20°C/-80°C, and the lyophilized form has a shelf life of 12 months at -20°C/-80°C.

Storage Condition

Store at -20°C/-80°C upon receipt. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.

Tag Info

The tag type is determined during manufacturing. If you require a specific tag, please inform us, and we will prioritize its development.

Synonyms

RpII15; CG3284; DNA-directed RNA polymerase II subunit RPB9; RNA polymerase II subunit B9; DNA-directed RNA polymerase II 15.1 kDa polypeptide

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-129

Protein Length

full length protein

Purity

>85% (SDS-PAGE)

Species

Drosophila melanogaster (Fruit fly)

Target Names

RpII15

Target Protein Sequence

MTTAFDAAHT EGPGFVGIRF CQECNNMLYP KEDKENKILL YACRNCDYKQ EADSNCIYVN KIMHEIDELT HIVPDVISDP TLPRTEDHAC PKCSHREAVF FQAQTRRAEE EMRLYYVCTN QNCTHRWTE

Uniprot No.

P36958

Target Background

Function

DNA-dependent RNA polymerase transcribes DNA into RNA using the four ribonucleoside triphosphates. RNA polymerase II, which synthesizes mRNA precursors and many functional non-coding RNAs, contains this subunit. Pol II, the core of the basal RNA polymerase II transcription machinery, is composed of mobile elements. RPB9, located in the 'upper jaw' surrounding the central cleft, is believed to bind the incoming DNA template.

Database Links

KEGG: dme:Dmel_CG3284

STRING: 7227.FBpp0082384

Protein Families

Archaeal RpoM/eukaryotic RPA12/RPB9/RPC11 RNA polymerase family

Subcellular Location

Nucleus, nucleolus.

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Recombinant Drosophila melanogaster DNA-directed RNA polymerase II subunit RPB9 (RpII15)

Description

Functional Roles in Transcription

Recombinant Production and Applications

Table 2: Biochemical Properties of Recombinant RpII15

Phenotypic Consequences of RpII15 Deficiency

Evolutionary and Mechanistic Insights

Open Questions and Future Directions

Product Specs

Target Background

Quick Inquiry

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