Evidence map›Paper›PMID 38094251›Full record

ArticleBio-protocol2023

Expression and Purification of Recombinant Human Mitochondrial RNA Polymerase (POLRMT) and the Initiation Factors TFAM and TFB2M.

An H Hsieh, Sean D Reardon, Jubilee H Munozvilla-Cabellon, Jiayu Shen, Smita S Patel, Tatiana V Mishanina

Open access · diamondAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Bio-protocol, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
1.2field-weighted citation impact, top 20% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

4 citing papers in PubMed, 8 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 3 institutions in 1 country.

An H HsiehDepartment of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
Sean D ReardonDepartment of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
Jubilee H Munozvilla-CabellonDepartment of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
Jiayu ShenDepartment of Biochemistry and Molecular Biology, Rutgers University, Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Smita S PatelDepartment of Biochemistry and Molecular Biology, Rutgers University, Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Tatiana V MishaninaDepartment of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
University of California, San Diego · USJohnson University · USRutgers, The State University of New Jersey · US

Funding

Mechanistic studies of nucleic acid enzymes involved in DNA replication, transcription, and innate immunity.R35GM118086 · NIGMS · RUTGERS BIOMEDICAL AND HEALTH SCIENCES · PI SMITA S PATEL · 2016 to 2026
$8.9M
MOLECULAR BIOPHYSICS TRAINING PROGRAMT32GM008326 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI KOMIVES, ELIZABETH A. · 1989 to 2020
$7.5M
Training in Multi-Scale Analysis of Biological Structure and FunctionT32EB009380 · NIBIB · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI Andrew D. McCulloch, Padmini Rangamani · 2009 to 2026
$4.8M
Mechanisms of transcription and co-transcriptional gene expression processes in bacteria and human mitochondriaR35GM142785 · NIGMS · UNIVERSITY OF CALIFORNIA, SAN DIEGO · PI Tatiana Vladimirovna Mishanina · 2021 to 2026
$2.5M
NIBIB NIH HHS T32 EB009380NIGMS NIH HHS R35 GM118086NIGMS NIH HHS R35 GM142785NIGMS NIH HHS T32 GM008326
6 · The paper itself

Abstract

Human mitochondrial DNA (mtDNA) encodes several components of oxidative phosphorylation responsible for the bulk of cellular energy production. The mtDNA is transcribed by a dedicated human mitochondrial RNA polymerase (POLRMT) that is structurally distinct from its nuclear counterparts, instead closely resembling the single-subunit viral RNA polymerases (e.g., T7 RNA polymerase). The initiation of transcription by POLRMT is aided by two initiation factors: transcription factor A, mitochondrial (TFAM), and transcription factor B2, mitochondrial (TFB2M). Although many details of human mitochondrial transcription initiation have been elucidated with in vitro biochemical and structural studies, much remains to be addressed relating to the mechanism and regulation of transcription. Studies of such mechanisms require reliable, high-yield, and high-purity methods for protein production, and this protocol provides the level of detail and troubleshooting tips that are necessary for a novice to generate meaningful amounts of proteins for experimental work. The current protocol describes how to purify recombinant POLRMT, TFAM, and TFB2M from

Indexed as

Bacterial protein expressionHeparinMaltose binding protein (MBP) fusion protein purificationNi-NTAPOLRMTProtein purificationTEV proteaseTFAMTFB2M

Identifiers

PMID38094251
PMCPMC10714150
OpenAlexW4389324081

What OpenQuestion holds

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LicenceCC BY-NC
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.