| 1986 |
PCCA encodes the alpha subunit of propionyl-CoA carboxylase (PCC), a biotin-dependent enzyme. The alpha subunit contains the covalently bound biotin prosthetic group and the biotin-binding domain, as confirmed by the presence of the Ala-Met-Lys-Met sequence (biotin binding site conserved across biotin-dependent carboxylases) in PCCA cDNA clones. The PCCA gene was chromosomally mapped to chromosome 13 using somatic mouse-human hybrid panels. |
cDNA cloning, oligonucleotide probing, somatic cell hybrid panel mapping, Northern blot |
Proceedings of the National Academy of Sciences of the United States of America |
High |
3460076
|
| 1987 |
In pccA complementation group patients (PCCA-deficient), alpha-chain mRNA is absent in most fibroblast strains, while beta-chain mRNA is present. The beta subunit (PCCB) is rapidly degraded in the absence of the alpha subunit, indicating that alpha-beta subunit interaction is required for stabilization of the beta chain. This confirms that PCCA encodes the alpha subunit of PCC. |
Northern blot with PCCA and PCCB cDNA probes, isotope-tracer labeling and immunoprecipitation of fibroblast extracts |
American journal of human genetics |
High |
3687944
|
| 1993 |
Full-length PCCA cDNA, when expressed by DNA-mediated gene transfer in pccA-deficient fibroblasts, reconstitutes propionate flux to normal levels, confirming that PCCA alone is sufficient to rescue the enzymatic defect. Maximum PCC holoenzyme activity upon PCCA expression reached only 10-20% of normal controls (corresponding to transfected cell fraction), indicating PCCA expression level does not normally limit PCC holoenzyme activity or propionate flux. |
cDNA expression (gene transfer), propionate flux assay, PCC holoenzyme activity measurement in fibroblasts |
American journal of human genetics |
High |
8434582
|
| 1999 |
PCCA is a mitochondrial heteropolymeric enzyme composed of alpha and beta subunits (encoded by PCCA and PCCB genes, respectively), involved in catabolism of branched-chain amino acids, odd-chain fatty acids, cholesterol, and other metabolites. In PCCA patients, combined absence of both alpha and beta subunits is observed by Western blot, indicating that the alpha subunit is required for stability of the assembled complex. |
Western blot, complementation assay, Northern blot, RT-PCR of patient fibroblasts |
Human mutation |
High |
10502773
|
| 1999 |
The PCCA pre-mRNA contains a constitutive 84-bp cryptic (pseudo)exon derived from an intron between nucleotides 1209 and 1210. This cryptic exon is normally present at very low levels in all cells but becomes relatively detectable when the normal-length mRNA is destabilized by nonsense or frameshift mutations (R288X, 700del5, 1115del4, 1671IVS+5G->C). Incorporation of the 84-bp insertion causes translation termination via two in-frame stop codons. |
RT-PCR, sequencing, comparative analysis of patient and normal fibroblast cell lines |
Human molecular genetics |
High |
9887338
|
| 2001 |
The PCCA gene spans more than 360 kb, consists of 24 exons (37–335 bp each), and the translation initiation codon is located 75 nucleotides upstream of the previously accepted site. The 5'-flanking region contains a putative CpG island (extending into exon 1 and part of intron 1), lacks a TATA box, but contains AP-1 sites and a consensus Sp1 (GC box) binding sequence in the proximal promoter. |
EST analysis, RT-PCR, genomic sequencing, promoter sequence analysis |
Molecular genetics and metabolism |
Medium |
11592820
|
| 2002 |
Eleven PCCA missense mutations and one in-frame deletion, when expressed in patient fibroblasts and in a cell-free in vitro system, result in reduced PCC enzyme activity and increased protein turnover/instability. Most mutant proteins show an increased rate of degradation, indicating structural alterations incompatible with normal assembly into a stable, functional PCC oligomer. |
Expression in deficient fibroblasts, cell-free in vitro expression, PCC activity assay, protein stability assessment |
Biochimica et biophysica acta |
High |
12385775
|
| 2009 |
Large genomic deletions in the PCCA gene (including frequent deletions of exons 3-4 and exon 23) cause propionic acidemia. A deletion of exons 3 and 4 results in an in-frame deletion of 39 amino acids; this in-frame deletion was expressed in a eukaryotic system and confirmed as pathogenic (loss of PCC activity). The high frequency of large deletions may be due to the abundance of intronic repetitive elements in the PCCA gene. |
MLPA, long-PCR, eukaryotic expression system, PCC activity assay |
Molecular genetics and metabolism |
Medium |
19157943
|
| 2017 |
In C. elegans, deletion of pcca-1 (ortholog of human PCCA) globally impairs mitochondrial energy metabolism: reduces mitochondrial oxidative phosphorylation capacity and efficiency, increases mitochondrial matrix oxidant burden, decreases mitochondrial membrane potential and content, and inhibits distal TCA cycle flux. These findings indicate that PCC/PCCA deficiency causes broader metabolic dysfunction beyond toxic propionyl-CoA precursor accumulation. |
C. elegans gene deletion, direct polarography of isolated mitochondria, in vivo mitochondrial physiology quantitation, UPLC amino acid profiling, GC/MS with 13C-glucose metabolic flux analysis |
Journal of inherited metabolic disease |
High |
29159707
|
| 2021 |
Twenty-four variants in PCCA (and PCCB) genes predicted to affect splicing were tested by minigene splicing assay; 13 variants (including one missense and two synonymous variants) caused significant alteration of splicing with predicted loss-of-function at the protein level, confirming their pathogenic mechanism via aberrant PCCA pre-mRNA splicing. |
Minigene splicing assay, RT-PCR, sequencing |
International journal of molecular sciences |
Medium |
33923806
|
| 2024 |
PCCA (mitochondrial carboxylase) physically interacts with and colocalizes with Listeria monocytogenes phospholipase PlcB within host cells. The amino acids 504–508 of PCCA are critical for this interaction. Overexpression of PCCA (via pCMV-N-HA-PCCA plasmid) reduces L. monocytogenes proliferation, while siRNA knockdown of PCCA increases bacterial proliferation, demonstrating an inverse correlation between PCCA levels and bacterial survival. L. monocytogenes infection does not significantly alter PCCA expression levels. |
Co-immunoprecipitation/colocalization, siRNA knockdown, plasmid overexpression, bacterial proliferation assay, mRNA/protein expression analysis in HeLa cells |
Applied and environmental microbiology |
Medium |
38727222
|
| 2024 |
PPDPF (pancreatic progenitor cell differentiation and proliferation factor) interacts with PCCA (identified by mass spectrometry) and blocks the interaction between PCCA and its partner subunit PCCB, thereby inhibiting PCC-dependent methionine catabolism via the C-Vomit pathway. This leads to elevated intracellular methionine and S-adenosylmethionine (SAM) levels and promotes esophageal squamous cell carcinoma progression. |
Mass spectrometry interaction screen, co-immunoprecipitation, metabolite measurement (methionine, SAM), PPDPF knockdown in vivo and in vitro |
Cancer letters |
Medium |
39694223
|
| 2025 |
lncBADR binds directly to PCCA (and Mccc1) in T cells, inhibiting BCAA degradation. Knockout of lncBADR in T cells restores PCCA-mediated BCAA catabolism, decreasing intracellular BCAA levels and reducing mTOR-Stat1 signaling and IFN-γ secretion. High-BCAA feeding partially reversed the protective effects of lncBADR knockout, confirming that lncBADR acts through PCCA-dependent BCAA metabolism. |
T cell-specific lncBADR knockout mice, RNA binding assays, metabolite profiling, mTOR-Stat1 pathway analysis, IFN-γ measurement, rescue experiment (high-BCAA feeding) |
Journal of neuroinflammation |
Medium |
41013574
|
| 2024 |
In Pcca-/-(A138T) mice (a propionic acidemia model), fasting reduces propionylcarnitine, the C3/C2 ratio, ammonia, and methylcitrate. This is attributed to significant reduction in microbiome-produced propionate and increased fatty acid oxidation (decreasing propionyl-CoA synthesis and enhancing acetyl-CoA synthesis) during fasting. Fasting-induced gluconeogenesis further facilitates propionyl-CoA catabolism without changing propionyl-CoA carboxylase activity. |
Pcca mutant mouse model, metabolite profiling (propionylcarnitine, C3/C2, ammonia, methylcitrate), measurement of PCC enzymatic activity, assessment of microbiome-derived propionate |
Communications biology |
Medium |
38811689
|
| 2026 |
A deep-intronic PCCA variant (c.1285-1358C>G) causes constitutive inclusion of the known 84-bp pseudoexon in PCCA mRNA, abolishing functional PCCA and PCCB protein expression and severely reducing PCC activity. Antisense oligonucleotides (ASOs) targeting this pseudoexon restore productive PCCA splicing, rescue PCCA protein expression, and markedly increase PCC activity above wild-type levels in patient fibroblasts. ASO treatment was also effective in other PA fibroblast lines with residual activity >1% of normal. |
Patient fibroblast analysis, RT-PCR/sequencing, ASO transfection, PCC enzymatic activity assay, protein expression analysis |
Molecular therapy. Nucleic acids |
High |
42028575
|