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Articles

Biotin Regulates the Genetic Expression of Holocarboxylase Synthetase and Mitochondrial Carboxylases in Rats¹

Unidad de Genética de la Nutrición of the Instituto de Investigaciones Biomédicas, UNAM and Instituto Nacional de Pediatría, México DF 04530

²To whom correspondence should be addressed. E-mail: velare{at}servidor.unam.mx

	ABSTRACT

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Biotin is the cofactor of carboxylases [pyruvate (PC), propionyl-CoA (PCC), 3-methyl crotonyl-CoA and acetyl-CoA], to which it is covalently bound by the action of holocarboxylase synthetase (HCS). We have studied whether biotin also regulates their expression, as it does other, nonrelated enzymes (e.g., glucokinase, phosphoenol pyruvate carboxykinase, guanylate cyclase). For this purpose, HCS, PC and PCC mRNAs were studied in biotin-deficient rat liver, kidney, muscle and brain of biotin-deficient rats. PC- and PCC-specific activities and protein masses were also measured. The 24-h time course of HCS mRNA in deficient rats was examined after biotin supplementation. HCS mRNA was significantly reduced during vitamin deficiency. It increased in deficient rats after biotin was injected, reaching control levels 24 h after administration. These changes seem to be the first known instance in mammals of an effect of a water-soluble vitamin on a mRNA functionally related to it. In contrast, the decreased activities of the carboxylases were associated with reductions in the amounts of their enzyme proteins except in brain. However, their mRNA levels were not affected. There are no reports on these types of vitamin affecting the mRNA or protein levels of their apoenzymes or their products. This work provides evidence for biotin being a modulator of the genetic expression of the enzymes involved in its function as a cofactor. As such, it may be a useful model for probing a similar role for other water-soluble vitamins.

KEY WORDS: • biotin • carboxylases • holocarboxylase synthetase • gene expression • rats

	INTRODUCTION

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Besides their well-known roles as substrates and cofactors, nutrients have recently been found to function as regulators of gene expression (1) . Among them are polyunsaturated fatty acids (2) , cholesterol (3 ,4) , glucose and fructose (5) , specific minerals such as iron (6) and lipid-soluble vitamins like retinoic acid (7 ,8) . As such, they act at different levels, including nuclear proteins and cis-acting elements (9 ,10) . In contrast, little is known about water-soluble vitamins as genetic modulators (11 ,12) . In particular, we are not aware of any report of these vitamins regulating their coenzyme role through the mRNA of the enzyme responsible for the holoenzyme formation.

Biotin is the cofactor of the carboxylases of pyruvate (PC),³ propionyl CoA (PCC), 3-methyl crotonyl CoA (MCC) and acetyl CoA, the first three located in the mitochondria and the last in the cytosol. These enzymes participate in the metabolism of carbohydrates, lipids and proteins, catalyzing the carboxylation of different metabolites (13 14 15) . They are synthesized as inactive apocarboxylases. Holocarboxylase synthetase (HCS) catalyzes their activation by covalently binding biotin to a lysine residue (16) . In this article we present evidence that biotin regulates the genetic expression of the enzymes to which it is functionally related, at the mRNA level in the case of HCS and at the protein levels of carboxylases (PC and PCC). The effects of biotin deficiency were studied in rat liver, kidney, muscle and brain.

	MATERIALS AND METHODS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Reagents and chemicals.

[-³²P] deoxycytidine triphosphate [dCTP (30 Ci/mmol)], sodium [¹⁴C] bicarbonate (58.0 mCi/mmol) and Megaprime DNA labeling system were purchased from Amersham Pharmacia Biotech (Piscataway, NJ). TRIzol reagent, oligo (dt) cellulose columns, M-MLV reverse transcriptase (40,000 U) kit and 2'-deoxynucleoside 5'-triphosphate were purchased from Life Technologies (Gaithersburg, MD). Amplificase (5 U/µL) was purchased from Biotecnologías Universitarias (México City, Mexico). Geneclean II kit was purchased from BIO 101. ATP, d-biotin, acetyl-CoA, propionyl-CoA, glutathione and pyruvic acid were purchased from Sigma Chemicals (México City, Mexico). Nitroblue tetrazolium chloride, 5-bromo-4-chloro-3-indolyl phosphate toluidine salt, streptavidin-alkaline phosphatase conjugate and biotinylated sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis high range standards were purchased from Bio-Rad (México City, Mexico). Complete protease inhibitors were purchased from Roche Molecular Biochemicals (México City, Mexico).

Animals and biotin-deficient diet.

Male Wistar rats, ages 21 to 28 d (40–60 g), obtained from the Experimental Research Department at the National Institute of Pediatrics, México D.F were placed in air-filtered cages and were fed a biotin-deficient diet (17) (ICN Nutritional Biochemicals, Cleveland, OH). Environmental conditions and light-dark cycles (12:12 h) were strictly controlled. For each of the deficient rats, one control was injected with 200 µg of biotin in 1.0 mol/L phosphate buffer saline [PBS (pH = 7.0)] intraperitoneally. Rats were killed by intraperitoneal injection of 200 µL sodium pentobarbital (Pfizer, Mexico City, Mexico), at wk 8–10 of the experimental period, when manifestations of deficiency began to appear (see Results). Liver, kidney, abdominal muscle and brain were removed and frozen at -70°C. The protocols used in these experiments were approved by the Animal Care Committee of the National Institute of Pediatrics, México D.F.

Tissue homogenates.

The organs were thawed, washed with PBS and homogenized with a polytron (Kinematica AG, Littau, Switzerland), at 4°C for two pulses of 10 s each, with a 30-s pause between them, in three volumes of the homogenate buffer, pH 7.5 (0.5 mol/L, Tris HCl, 1 mol/L KCl, 10 mmol/L EDTA) containing the protease inhibitors mix. The homogenates were then sonicated (Branson cell disruptor 200, Danbury, CT) with five sonication pulses of 10 s each, with 1-min pauses between them, followed by centrifugation at 105,000 x g at 2°C for 10 min. The fat layer was discarded, and samples were obtained for protein determination by the Lowry method and for storage at -70°C.

Rat HCS cDNA cloning.

A segment of HCS cDNA was cloned from normal rat liver poly A RNA, using as primers oligonucleotides whose sequence was conserved among different distant species. For this purpose, degenerate oligonucleotides were designed aligning HCS yeast, mouse and human sequences to find conserved regions. The Codon Usage Table (CUT; http://www.dna.affrc.go.jp/nakamura-bin/showcodon.cgi.species=Rattus+norvegicus+[gbrod]) was used to obtain the least degenerate sequence to be used as primers: Sense 5'-AAG TGG CCC AAC GAY(C/T) ATT TAY 3'; antisense 5'-GTC GAA GGA GTT GCC GTC CGG 3'. Reverse transcription (RT) and polymerase chain reaction (PCR) (18) (GeneAmp PCR system; Perkin Elmer, Norwalk, CT) were used to achieve a rat cDNA fragment of HCS, using the oligonucleotides above mentioned. The reaction mix for RT contained total RNA 2 µg/2.5 µL, 5x buffer, dithiothreitol 4 mmol/L, dATP, dCTP, thymidine 5'-triphosphate and deoxyguanosine triphosphate 200 µmol/L, oligo d(T) and M-MLV reverse transcriptase 40 U, in 10 µL final volume. The reaction mix for PCR contained the primers 0.5 mmol/L each, buffer 10x, MgCl₂ 1.5 mmol/L, dATP, dCTP, thymidine 5'-triphosphate and deoxyguanosine triphosphate 200 µmol/L each, Taq polymerase 4 U in a final volume of 50 µL. The RT-PCR products were electrophoresed in a 1.5% agarose gel and purified with Geneclean II kit. Once the rat HCS cDNA fragment was obtained, it was sequenced (Fig. 1 ), and new oligonucleotides were designed from the terminal regions of this fragment. The final sequences of the specific primers were: Sense 5' TAC AGC CTC TAT GAA GAT CG 3', Antisense 5' CTC CAC ACC CCA CCA CC 3'; and subsequently used for Southern blot analysis.

View larger version (48K): [in this window] [in a new window]   Figure 1. Nucleotide and derived amino acid sequence from a rat HCS cDNA fragment. A segment of HCS cDNA was cloned from normal rat liver poly A RNA, using as primers oligonucleotides whose sequence was conserved among different distant species. This fragment corresponds to the human HCS sequences between region between amino acid 570 and 717. The sequences of the specific primers utilized for semiquantitative determination of HCS mRNA, by RT-PCR analysis, are shown in bold type.

Total RNA was obtained from the different organs of five control and five deficient rats. Total RNA was isolated using TRIzol reagent (19) . Its concentration was determined by absorbance at 260 nm and its integrity was verified by electrophoresis on 1.1% denaturing agarose gels in the presence of 2.2 mol/L formaldehyde. Total RNA (4 µg) was reverse transcribed to synthesize single-strand cDNA. The RT reaction mixture (10 µL) was subjected to PCR to amplify the HCS cDNA fragment, as described above. The other 10 µL of the cDNA was used to amplify simultaneously a fragment of the rat actin gene, which was used as a constitutive expression control (sense 5'-GGG TCA GAA GGA TTC CTA TG 3', antisense 5'-GGT CTC AAA CAT GAT CTG GG 3'). The 50 µL PCR included 10 µL of previously synthesized cDNA, 20 mmol/L Tris-HCl (pH 8.3), 50 mmol/L KCl, 1 mmol/L MgCl₂, 200µmol/L of each 2'-deoxynucleoside 5'-triphosphate, 0.5µmol/L of each primer and 2.5 U of Taq DNA polymerase. Negative controls without RNA and with nonretrotranscribed RNA were included in all the experiments. After an initial denaturation step at 95^°C for 5 min, the PCR was performed 30 cycles for HCS. The cycle profile for HCS and actin gene amplification was 95^°C, 1 min; 50^°C, 1 min; 72^°C, 1 min and a final extension was performed at 72^°C, 5 min. The number of cycles performed was within the exponential phase of the amplification process previously determined. PCR products were separated on 1.5% agarose gel, stained with ethidium bromide and subjected to Southern blot analysis. The gels were transferred to GeneScreen membranes and hybridized with radiolabeled probe. Probes (for HCS and actin) were radiolabeled with -³²P dCTP (20 µCi). After hybridization, membranes were washed twice with 2x SSC (sodium chloride/sodium citrate) at room temperature for 15 min. The membranes were exposed to Hyperfilm ßMax films for 24 h at -70°C. The closely approximated levels of HCS mRNA were estimated from the intensity of the bands in the autoradiographs, quantified by densitometry (see below). The data were normalized with the estimated mRNA actin levels, similarly obtained. Statistical analysis was performed using Student’s t test.

Time-course effect of biotin on HCS mRNA.

Six deficient biotin rats were injected intraperitoneally with 200 µg of biotin in 1.0 mol/L PBS (pH 7.0) and killed at different times: 0, 30 min, 1 h, 2 h, 4 h and 24 h. Each liver was obtained, stored at -70°C and processed separately. RNA total was extracted using TRIzol reagent. RT-PCR analysis was made as described above.

Carboxylase assays.

The homogenates were thawed, resuspended in 200 µL of lysis buffer and sonicated twice on ice (10 s each) with a 20-s pause between each sonication. They were then centrifuged at 750 x g at 2°C for 10 min, and the supernatant was used. PCC and PC activities were determined using the assays by Burry et al. (20) and the enzyme activities were expressed as nmol CO₂ fixed · min^-1 · mg protein^-1.

Carboxylase mass determination.

The mass of the carboxylase was estimated by streptavidin blots after in vitro biotinylation of tissue homogenates, to convert apo to holocarboxylases. Homogenate apocarboxylases were biotinylated by a technique modified from Desjardins and Dakshinamurti (21) . Aliquots of the homogenates containing 500 µg protein were added to the reaction mix containing 60 mmol/L Tris-HCl (pH 7.5), 0.82 mmol/L biotin, 0.1 mmol/L EDTA, bovine serum albumin 0.60 g/L, 3 mmol/L reduced glutathione, 8mmol/L MgCl₂ and 10 mmol/L ATP) in a final volume of 500 µL. It was incubated at 37°C for 6 h; additional ATP-MgCl₂ mix (26.75 µL) was added at the h 2 and 4 of incubation (22) . The reaction was stopped by the addition of 500 µL of electrophoresis buffer [0.5 mol/L Tris-HCl (pH 6.8), 40% glycerol, 10% SDS and 0.5% bromophenol blue) and was stored at -20°C. Maximal homogenate biotinylation was obtained by optimizing the reagents concentrations and the incubation time. Nonbiotinylated controls were similarly incubated, except that no biotin and ATP were added to the reaction mix. Aliquots containing 5 µg of homogenate protein were added to a SDS 8% polyacrylamide gel and electrophoresed (SDS-polyacrylamide gel electrophoresis) at 70 mA for 1 h. The gel was incubated with transfer buffer (48 mmol/L Tris, 39 mmol/L glycine, 1.3 mmol/L SDS, 20% methanol) with slow agitation for 30 min and blotted on a nitrocellulose membrane, 0.45 µm (Bio-Rad, Hercules, CA), using a semidry transfer cell (Bio-Rad), at 17 V for 30 min. The membrane was incubated with streptavidin-alkaline phosphatase to detect biotinylated proteins (23) . It was soaked in 50 mL of 5% blotto buffer (100 mmol/L boric acid, 47 mmol/L sodium borate, 75 mmol/L NaCl, 50 g/L powder milk) at 4°C, with slow agitation for 1 h, and washed with 80 mL of replicate buffer [90 g/L NaCl, 10 mmol/L Tris-HCl (pH 7.4), 0.50% Tween 20]. It was then incubated in 50 mL of wash buffer (0.15 mol/L NaCl, 0.25% Triton X 100, 20 mmol/L NaH₂PO₄) containing 10 µL streptavidin-alkaline phosphatase, at room temperature for 2 h. Afterward, it was washed three times with replicate buffer for 5 min each time and with 0.15 mol/L Tris-HCl (pH 8.8). Color development was performed incubating the membrane in the dark, with slow agitation, in 100 mL A-P buffer [10 mmol/L Tris base (pH 9.5), 10 mmol/L NaCl, 5 mmol/L MgCl₂], containing 50 µL nitroblue tetrazolium chloride solution (nitro blue tetrazolium 0.3 g in 1 mL 70% N'N-dimethylformamide) and 50 µL 5-bromo-4-chloro-3-indolyl phosphate toluidine salt solution (5-bromo-4-chloro-3-indolyl phosphate 0.15 g in 1 mL 70% dimethylformamide). Once the bands became visible, the membrane was washed with double-distilled water. Under these electrophoretic conditions, there was no clear separation of PCC and MCC.

Semiquantitative analysis of PC and PCC mRNA.

The mRNA levels of PC and PCC were determined by coamplification with RT-PCR method, with the following modifications. The amplification primers for PC were sense 5'-ACT TGT ATG AGC GGG ACT GC 3', antisense 5'-TGA CCT TGA CGG GGA TTG GA 3') (13) and for PCC were sense 5'-GA TGC CAG CTC GGT TCA TGT 3', antisense 5'-GAG GCC TTG ATC ATC ACA GG 3' (24) . The cycle profile was also similar to HCS and actin gene amplification, except 60°C, 1 min for annealing. Everything else was the same, including the determination of the number of cycles under which the amplification was exponential.

Densitometric analysis.

The electrophoretic bands obtained for Northern and streptavidin blot analyses were digitalized using a Scan Jet 3C (Hewlett-Packard, Palo Alto, CA) and Deskscan software (Hewlett-Packard), and the intensity of the bands was estimated by Collage software (Hewlett-Packard).

	RESULTS

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Determination of biotin nutritional status.

The rats fed the biotin-deficient diet, but not the controls, exhibited clear deficiency signs after wk 8 of treatment, including low body weight, hair loss, conjunctivitis and periorificial skin rash. The carboxylase activities were reduced (see below) and there was an abnormally high urinary excretion of 3-hydroxyisovaleric acid (data not shown), a marker of biotin deficiency (25 ,26) .

Holocarboxylase synthetase mRNA levels in biotin deficiency.

HCS mRNA levels were significantly lower in the four organs of the biotin-deficient rats than in controls (Table 1 ). These differences were more marked in liver and kidney than in muscle and brain (Fig. 2 ); the mRNA in these organs of deficient rats being less than one-half that of controls. When biotin was injected into deficient rats, the HCS mRNA liver increased to near normal levels. However, this was a delayed effect, apparent at 24 h after administration but not during the first few hours (Fig. 3 ).

View larger version (54K): [in this window] [in a new window]   Figure 2. RT-PCR analysis of HCS mRNA levels from liver, kidney, muscle and brain of control and biotin-deficient rats. RNA was reverse-transcribed and amplified by PCR using specific primers for rat HCS and actin, like constitutive control. PCR products were separated on 2% agarose gel and subjected to Southern blot analysis. C, control; D, biotin-deficient.

View larger version (41K): [in this window] [in a new window]   Figure 3. Time-course effect of biotin on HCS mRNA levels in liver from biotin-deficient rats. Six biotin-deficient rats were injected with biotin (200 µg) and each killed at a different time: 0, 30 min, 1 h, 2 h, 4 h and 24 h. The livers were processed as mentioned in Figure 1C , control.

Pyruvate and propionyl CoA carboxylase activities were significantly reduced in the organs of the deficient rats (Figs. 4 and 5 ). These differences generally were associated with significantly lower amounts of their enzyme proteins. In brain, the PC and PCC mass did not differ between groups, and their decrements in activity were fairly moderate. PC and PCC mRNA levels did not differ between groups in any organ studied.

View larger version (34K): [in this window] [in a new window]   Figure 4. Effects of biotin deficiency on activity, carboxylase mass and mRNA levels of the PC in organs of male Wistar rats fed a biotin-deficient diet or control and killed by intraperitoneal injection of 200 µL of sodium pentobarbital. *P < 0.05, **P < 0.01 and ***P < 0.005 vs. control. ND, not detected.

View larger version (37K): [in this window] [in a new window]   Figure 5. Effects of biotin-deficiency on activity, carboxylase mass and mRNA levels of the PCC in organs of male Wistar rats fed a biotin-deficient diet or control and killed by intraperitoneal injection of 200 µL of sodium pentobarbital. Protein (middle panel) includes MCC because it comigrates with PCC. *P < 0.05, **P < 0.01 and ***P < 0.005 vs. control

	DISCUSSION

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Regulation of genetic expression by vitamins has been more comprehensively studied for the lipid (8 ,28 ,29) than for the water-soluble vitamins (11 ,12 ,30 ,31) . The HCS mRNA changes reported here seem to be the first known instance in mammals of an effect of a water-soluble vitamin on the mRNA of an enzyme that acts directly with that vitamin. The observed changes were quite evident, reproducible and significant. In contrast, the apparent variation in the HCS mRNA amounts among organs likely lacks biological importance, because the densitometric conditions used to observe the bands varied from organ to organ. Information on possible differences in the amount or activity of its protein product is not currently available.

Small molecules may have a direct effect on mRNA levels, e.g., by interacting with transcription factors or with RNA-binding proteins (32) . Biotin has been shown to be present in the cell nucleus together with a 60-kDa biotin-binding protein, to which this vitamin binds reversibly in vitro (33) , suggesting the possibility of gene regulation through a nuclear receptor, as with vitamins A and D (8 ,34) . However, the long lapse between biotin administration to deficient rats and the recovery of the HCS mRNA levels suggests a more indirect action, requiring metabolic processing, e.g., protein synthesis, or affecting endocrine or signaling pathways. In this respect, it is interesting that increased levels of guanosine 3',5'-cyclic monophosphate, mediated by guanylate cyclase (35) , were associated with biotin-induced expression of hepatic glucokinase (36 ,37) and the asialoglycoprotein receptor (38) . These results suggest that second messenger (guanosine 3',5'-cyclic monophosphate) may be involved in the effects evoked by biotin. Additional studies are needed to clarify the mechanism(s) involved in the HCS mRNA changes reported herein.

Along with the decrease of HCS mRNA, not only the activity, but also the actual amounts (mass) of carboxylases (PC and PCC) were reduced in the biotin-deficient rats, although their mRNA levels were unaffected. ApoPC and apoPCC, devoid of their cofactor, might be more prone to degradation than their holo forms (39) . Alternatively, biotin deficiency may reduce the synthesis of PCC and PC (40) .

It is interesting that PC was unaffected in the brain of the biotin-deficient rats. It may be that relatively normal concentrations of biotin are maintained locally, because a similar result was observed for brain PCC, with a lesser reduction of the carboxylase activities, relative to other organs. This is supported by the observation that brain holoPC and holoPCC concentrations (measured in streptavidin blots of homogenates incubated without biotin and ATP; results not shown) were sustained in the deficient rats. It will be important to identify those steps in the metabolism of biotin in the brain, e.g., its transport across the blood-brain barrier (41) , which may contribute to spare this organ during vitamin deficiency.

In conclusion, this work provides evidence for biotin as modulator of the genetic expression of the enzymes involved in its function as a cofactor. As such, it may be a useful model for probing a similar role for other water-soluble vitamins.

	ACKNOWLEDGMENTS

 
We thank students Daniel Ortega, Merlin Toledo, Gabriela Sandoval and Victor Serrano. We also thank Alfonso León Del Rio and Ignacio Camacho Arroyo for their helpful advise and continued support. Finally, Margarita Terán, Silvestre Frenk and Stephen Cederbaum carefully read the manuscript and contributed many useful comments and suggestions.

	FOOTNOTES

 
¹ Supported by Research Grant 27973-M from the Consejo Nacional de Ciencia y Tecnología (CONACyT).

³ Abbreviations used: dCTP, deoxycytidine triphosphate; HCS, holocarboxylase synthetase; MCC, 3-methyl crotonyl CoA; PBS, phosphate buffer saline; PC, pyruvate carboxylase; PCC, propionyl-CoA carboxylase; PCR, polymerase chain reaction; RT, reverse transcription; SDS, sodium dodecyl sulfate.

Manuscript received January 12, 2001. Initial review completed February 15, 2001. Revision accepted April 9, 2001.

	REFERENCES

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 

1. Hanson R. W. Nutrient control of gene transcription minireview series. J. Biol. Chem. 2000;275:30747[Free Full Text]

2. Duplus E., Glorian M., Forest C. Fatty acid regulation of gene transcription. J. Biol. Chem. 2000;275:30749-30752[Free Full Text]

3. Dawson P. A., Metherall J. E., Ridgway N. D., Brown M. S., Goldstein J. L. Genetic distinction between sterol-mediated transcriptional and posttranscriptional control of 3-hydroxy-3-methylglutaryl-coenzyme A reductase. J. Biol. Chem. 1991;266:9128-9134[Abstract/Free Full Text]

4. Osborne T. F. Sterol regulatory element-binding proteins (SREBPs): key regulators of nutritional homeostasis and insulin action. J. Biol. Chem. 2000;275:32379-32382[Free Full Text]

5. Vaulont S., Vasseur-Cognet M., Kahn A. Glucose regulation of gene transcription. J. Biol. Chem. 2000;275:31555-31558[Free Full Text]

6. Klausner R. D., Harford J. B. Cis-trans models for post-transcriptional gene regulation. Science 1989;246:870-872[Free Full Text]

7. Fernandez-Mejia C., Davidson M. B. Regulation of glucokinase and proinsulin gene expression and insulin secretion in RIN-m5F cells by dexamethasone, retinoic acid, and thyroid hormone. Endocrinology 1992;130:1660-1668[Abstract]

8. Nagpal S., Chandraratna R. A. Vitamin A and regulation of gene expression. Curr. Opin. Clin. Nutr. Metab. Care 1998;1:341-346[Medline]

9. Clarke S. D. Nutrient regulation of gene expression: a methodological strategy. Miner. Electrolyte Metab. 1997;23:130-134[Medline]

10. Clarke S. D., Abraham S. Gene expression: nutrient control of pre- and posttranscriptional events. FASEB J 1992;6:3146-3152[Abstract]

11. Nagao M., Tanaka K. FAD-dependent regulation of transcription, translation, post-translational processing, and post-processing stability of various mitochondrial acyl-CoA dehydrogenases and of electron transfer flavoprotein and the site of holoenzyme formation. J. Biol. Chem. 1992;267:17925-17932[Abstract/Free Full Text]

12. Gulati S., Brody L. C., Banerjee R. Posttranscriptional regulation of mammalian methionine synthase by B12. Biochem. Biophys. Res. Commun. 1999;259:436-442[Medline]

13. Jitrapakdee S., Booker G. W., Cassady A. I., Wallace J. C. The rat pyruvate carboxylase gene structure: alternate promoters generate multiple transcripts with the 5'-end heterogeneity. J. Biol. Chem. 1997;272:20522-20530[Abstract/Free Full Text]

14. Moss J., Lane M. D. The biotin-dependent enzymes. Adv. Enzymol. 1971;35:321-398

15. Wolf B. Disorders of biotin metabolism. Scriver C. R. Beaudet A. L. Sly W. S. Valle D. eds. The Metabolic and Molecular Bases of Inherited Disease 1995:3151-3177 McGraw-Hill New York, NY.

16. León-del-Río A., LeClerc D., Ackerman B., Wakamatsu N., Gravel R. A. Isolation of a cDNA encoding human holocarboxylase synthetase by functional complementation of a biotin auxotroph of Escherichia coli. Proc. Natl. Acad. Sci. USA 1995;92:4626-4630[Abstract/Free Full Text]

17. Maeda Y., Kawata S., Inui Y., Fukuda K., Igura T., Matsuzawa Y. Biotin deficiency decreases ornithine transcarbamylase activity and mRNA in rat liver. J. Nutr. 1996;126:61-66

18. Camacho-Arroyo I., Pasapera A., Cerbón M. A. Regulation of progesterone receptor gene expression by sex steroid hormones in the hypothalamus and the cerebral cortex of the rabbit. Neurosci. Lett. 1996;214:25-28[Medline]

19. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 1987;162:156-159[Medline]

20. Burry B. J., Sweetman L., Nyhan W. L. Heterogeneity of holocarboxylase synthetase in patients with biotin responsive multiple carboxylase deficiency. Am. J. Hum. Genet. 1985;34:326-337

21. Desjardins P. R., Dakshinamurti K. Acetyl-CoA holocarboxylase synthesis in biotin-deficiency rat adipose tissue. Arch. Biochem. Biophys. 1971;142:292-298[Medline]

22. Kosow D. P., Lane M. D. Propionyl apocarboxylase activation catalyzed by cell-free enzyme extracts. Biochem. Biophys. Res. Commun. 1961;5:191-195[Medline]

23. Praul A. C., Brubaker K. O., Leach R. M., Gay C. V. Detection of endogenous biotin-containing proteins in bone and cartilage cells with streptavidin systems. Biochem. Biophys. Res. Commun. 1998;247:312-314[Medline]

24. Browner M. F., Taroni F., Sztul E., Rosenberg L. E. Sequence analysis, biogenesis, and mitochondrial import of the -subunit of rat liver propionyl-CoA carboxylase. J. Biol. Chem. 1989;264:12680-12685[Abstract/Free Full Text]

25. Mock N. I., Malik M. I., Stumbo P. J., Bishop W. P., Mock D. M. Increased urinary excretion of 3-hydroxyisovaleric acid and decreased urinary excretion of biotin are sensitive early indicators of decreased biotin status in experimental biotin deficiency. Am. J. Clin. Nutr. 1997;65:951-958[Abstract/Free Full Text]

26. Rodriguez-Melendez R., Perez-Andrade M. E., Diaz A., Deolarte A., Camacho-Arroyo I., Cicerón I., Ibarra I., Velázquez A. Differential effects of biotin deficiency and replenishment on rat liver pyruvate and propionyl-CoA carboxylases and on their mRNAs. Mol. Genet. Metab. 1999;66:16-23[Medline]

27. Chang H. I., Cohen N. D. Regulation and intracellular localization of the biotin holocarboxylase synthetase of 3T3–L1 cells. Arch. Biochem. Biophys. 1983;225:237-247[Medline]

28. Azzi A., Boscoboinik D., Clement S., Ozer N., Ricciarelli R., Stocker A. Vitamin E mediated response of smooth muscle cell to oxidant stress. Diabetes Res. Clin. Pract. 1999;45:191-198[Medline]

29. Chu F. F., Esworthy R. S., Lee L., Wilczynski S. Retinoic acid induces Gpx2 gene expression in MCF-7 human breast cancer cells. J. Nutr. 1999;129:1846-1854[Abstract/Free Full Text]

30. Allgood V. E., Cidlowski J. A. Vitamin B6 modulates transcriptional activation by multiple members of the steroid hormone superfamily. J. Biol. Chem. 1992;266:3819-3824

31. Allgood V. E., Powell-Oliver F. E., Cidlowski J. A. Vitamin B6 influences glucocorticoid receptor-dependent gene expression. J. Biol. Chem. 1990;265:12424-12433[Abstract/Free Full Text]

32. Hesketh J. E., Vasconcelos M. H., Bermano G. Regulatory signals in messenger RNA: determinants of nutrient-gene interaction and metabolic compartmentation. Br. J. Nutr. 1998;80:307-321[Medline]

33. Dakshinamurti K. C. Biotin-binding proteins. Dakshinamurti K. eds. Vitamin Receptors 1994:200-249 Cambridge University Press Cambridge, UK.

34. Carlberg C. Lipid soluble vitamins in gene regulation. Biofactors 1999;10:91-97[Medline]

35. Singh I., Dakshinamurti K. Stimulation of guanylate cyclase and RNA polymerase II activities in HeLa cells and fibroblasts by biotin. Mol. Cell. Biochem. 1988;79:47-55[Medline]

36. Chauhan J., Dakshinamurti K. Transcriptional regulation of the glucokinase gene by biotin in starved rats. J. Biol. Chem. 1991;266:1035-1038

37. Romero-Navarro G., Cabrera-Valladares G., German M. S., Matschinsky F. M., Velázquez A., Wang J., Fernández-Mejía C. Biotin regulation of pancreatic glucokinase and insulin in primary cultured rat islets and in biotin-deficient rats. Endocrinology 1999;140:4595-4600[Abstract/Free Full Text]

38. Collins J. C., Paietta E., Green R., Morell A. G., Stockert R. J. Biotin-dependent expression of the asialoglycoprotein receptor in HepG2. J. Biol. Chem. 1988;263:11280-11283[Abstract/Free Full Text]

39. Schimke R. T., Sweeeny E. W., Berlin C. M. The roles of synthesis and degradation in the control of rat liver tryptophan pyrrolase. J. Biol. Chem. 1965;240:322-331[Free Full Text]

40. Jitrapakdee S., Wallace J. Structure, function and regulation of pyruvate carboxylase. Biochem. J. 1999;340:1-16

41. Baur B., Baumgartner E. R. Biotin and biocytin uptake into cultured primary calf brain microvessel endothelial cells of the blood-brain barrier. Brain Res 2000;858:348-355[Medline]

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