Ch. 17-18 Flashcards
Gene Expression, Mitosis and Meiosis
gene expression
the process by which information encoded in DNA directs the synthesis of proteins, or RNAs that are not translated into proteins and instead function as RNAs
transcription
-the synthesis of RNA using a DNA template
-DNA is transcribed into RNA (same language)
-it starts and stops at specific sequences
translation
-the synthesis of a polypeptide using the genetic information encoded in an mRNA molecule
-RNA is translated into protein (different language; change of language from nucleotides to amino acids)
what bases does DNA use?
A, G, C, and T
what bases does RNA use?
A, G, C and U
(an RNA ‘U’ pairs with DNA ‘A’ during transcription)
what is the function of DNA in protein coding genes?
for protein coding genes, DNA serves as a template to produce a single strand messenger RNA (mRNA)
what does mRNA carry, and where to?
mRNA carries the genetic information to the ribosome
what occurs in the ribosome after mRNA carries the genetic information there?
the information is translated into proteins
how is transcription and translation different in bacteria?
in bacteria, transcription and translation are not separated into separate compartments, and they can occur simultaneously
where do eukaryotes export mRNA to and from, for translation?
eukaryotes must export the mRNA from the nucleus to cytoplasm for translation
pre-mRNA
-before the mRNA leaves the nucleus it starts out as pre-mRNA
-the pre-mRNA has certain regions removed, a cap is added to the 5’ end and additional ‘A’ nucleotides added to the 3’ end before it leaves the nucleus as a mature mRNA
primary transcript
the initial RNA transcript from any gene before it is processed; also applies to RNAs that are not translated into protein
template strand of DNA
-used to generate the mRNA
-during transcription the two strands of DNA separate, and only one of the two strands is used as the template for the mRNA
-for any gene, the same strand always serves as the template strand
-different genes on the same chromosome can use opposite strands of DNA as the template strand
-the mRNA is synthesized in the 5’ to 3’ direction, and it is antiparallel to the template strand
coding strand
-aka the nontemplate strand
-the nontemplate strand has the same nucleotide sequence as the mRNA, except that T is substituted for U
The Genetic Code
-there are only 4 bases in DNA and multiple nucleotides must be combined together to specify the different amino acids
triplet code
a genetic information system in which a set of three-nucleotide-long words specify the amino acids for polypeptide chains
codons
the mRNA nucleotide triplets
how was the genetic code determined?
by making synthetic mRNAs and combining them with ribosomes, amino acids, and other components in a test tube
-ex: an RNA molecule with only U (UUUUUUUUUUUU) would produce a polypeptide with only phenylalanine (PhePhePhePhe)
characteristics of the genetic code
-the genetic code is REDUNDANT, more than one codon is used for most amino acids
-the genetic code is NOT AMBIGUOUS, one codon codes for only one amino acid
AUG codon
-codes for methionine
-the start signal for translation
UAA, UAG, and UGA codons
-do not code for an amino acid
-they are the stop signals for translation
reading frame
-each mRNA will have three possible frames that can be translated into amino acids
-only one strand is used, called the reading frame
-generally begins from the first AUG in the mRNA sequence
the genetic code is universal
-the genetic code applies to all organisms
-same code used in bacteria, plants, and people
-implies that all life on earth had a common ancestor
-a useful feature for molecular biologists
RNA polymerase
-the enzyme that links ribonucleotides into a growing RNA chain during transcription, based on complementary binding to nucleotides on a DNA template strand
-does not need a primer, unlike DNA polymerase
-works in a 5’ to 3’ direction
-unwinds the DNA as it goes to expose the template strand
promoter
the site where RNA polymerase attaches and begins transcription
terminator
a specific sequence in bacteria that signals the end of transcription (termination is different in eukaryotes)
transcription unit
the stretch of DNA that is transcribed into RNA
the 3 phases of producing an RNA:
- initiation
- elongation
- termination
downstream
the direction of transcription (the termination sequence is downstream from the promoter)
upstream
the opposite direction of transcription
initiation of transcription
-in bacteria, RNA polymerase binds to a specific sequence in the promoter
transcription factors
-other proteins in eukaryotes that bind that bind to the DNA first
-typically have a DNA binding domain and a protein interaction domain
general transcription factors
-a protein that binds to DNA to initiate the transcription of genetic information into messenger RNA, for eukaryotes
-poly A signal sequence
sigma factor
-a protein in bacteria/prokaryotes that is necessary for the start of transcription
-GC rich terminator sequence
transcription initiation complex
when RNA polymerase forms a complex with the transcription factors
RNA polymerase II
used to transcribe mRNA (eukaryotes have three RNA polymerases)
TATA box
-the many promoters in eukaryotes that contain a specific sequence TATAAAA, to which transcription factors bind in order to establish the transcription initiation complex
Pribnow box
-a six-nucleotide sequence of TATAAT that is a vital part of the promoter site on DNA for transcription to occur in bacteria/prokaryotes
start point
-the site where transcription actually begins
-the promoter consists of DNA sequences dozens of nucleotides upstream from the start point
elongation of transcription
-RNA polymerase untwists the DNA, exposing about 10-20 nucleotides at a time
-RNA nucleotides that are complementary to the DNA template are added to the 3’ end of the growing RNA molecule
termination of transcription in bacteria
-the terminator sequence in the DNA is transcribed into RNA, and the newly formed RNA forms a structure that causes the polymerase to fall off the DNA
-the mRNA in bacteria doesn’t need to be processed and translation can begin
termination of transcription in eukaryotes
-the RNA polymerase passes through a specific sequence in the DNA that creates a polyadenylation signal (AAUAAA) in the pre-mRNA molecule
- 10-31 nucleotides downstream of the polyadenylation signal proteins that associate with the newly formed pre-mRNA cut it free from the RNA polymerase
extensive processing of pre-mRNA in eukaryotes
-before the pre-mRNA can leave the nucleus it undergoes extensive processing that alters both ends of the RNA and cuts sequences out of the middle
-the 5’ end of the pre-mRNA receives a 5’ cap, which is a modified guanine nucleotide
-a special enzyme adds 50 – 250 adenine nucleotides to the 3’ end. This long stretch of As is called a poly-A tail
-the 5’ cap and poly-A tail protect the mRNA from degradation, are used to export the mRNA form the nucleus, and help to attach ribosomes to the 5’ end of the RNA
Untranslated Regions (UTRs)
the regions of RNA that are not translated (not all of the RNA nucleotides will be translated into amino acids)
RNA splicing
-most eukaryotic genes and the RNA transcripts produced from them have long stretches of nucleotides that are not translated into protein
-the sequence of DNA nucleotides that codes for a polypeptide are not contiguous, but are split into regions
-the 5’ UTR and 3’ UTR are included in exons, even though they are not translated into proteins
introns
-the noncoding regions of nucleotides that lie between coding regions
-aka intervening sequences
exons
the regions of nucleotides that are expressed (usually translated into protein)
splicing
the process in which intervening sequences (introns) are cut out of the pre-mRNA (primary transcript)
splicesosome
-a large complex made up of proteins and RNA molecules that splices RNA by interacting with the ends of an RNA intron, releasing the intron, and joining the two adjacent exons
-contains several small nuclear ribonucleoproteins (snRNPs)
ribonucleoproteins (snRNPs)
each snRNP contains a small nuclear RNA (snRNA) that can act as a catalyst in the splicing process
ribozymes
-are RNA molecules that function as catalysts (therefore not all biological catalysts are made of protein)
-some organisms only use snRNAs to catalyze splicing without using any proteins
why can RNAs operate as enzymes?
-because they can adopt specific three dimensional shapes, and the bases contain functional groups that can interact with other molecules
what gives RNA a high degree of specificity?
the ability to form complementary bonds with other nucleic acids gives RNAs a high degree of specificity
alternative RNA splicing
-a process that creates different mRNA molecules from the same primary transcript, depending on which RNA segments are treated as exons and which as introns
-multiple proteins can be produced from the same gene (with alternative splicing different, but related proteins can be made from the same gene)
domains
-discrete structural and functional regions that proteins can have (ex: a DNA binding domain, or an active site for an enzyme, or a kinase domain)
-different exons can code for different domains
how can alternative splicing produce proteins with different functions?
by adding or removing specific domains
Prokaryotes versus Eukaryotes
-cellular location: cytoplasm (prok) vs nucleus (euk)
-AT rich promoter: Pribnow box (prok) vs TAT box (euk)
-proteins aid in RNA polymerase binding: sigma factor (prok) vs general transcription factors (euk)
-no mRNA modification, translation immediately occurs after mRNA synthesis (prok) vs extensive mRNA modifcation occurs prior to transport out of nucleus (euk)
what does translation bring together in order to make a protein?
mRNA, rRNA, and tRNA
mRNA
carries the genetic information from the DNA
rRNA molecules
are ribozymes that are integral parts of ribosomes where translation occurs
tRNA
converts the codons in the mRNA to the proper amino acid in the polypeptide
tRNA molecule
-a tRNA molecule is a short, single strand of RNA that adopts a specific shape, containing three loop domains
-one of the loops contains a triplet anticodon that is complementary to the codon in mRNA
-each tRNA molecule with a specific anticodon carries a specific amino acid
anticodon
complementary to the codon in mRNA
aminoacyl-tRNA synthetases
-link the correct amino acid to the correct tRNA
-there are 20 different aminoacyl-tRNA synthetases, one for each amino acid
active site of the aminoacyl-tRNA synthetase
-fits only a specific combination of amino acid and tRNA
-this molecular recognition ensures that the correct amino acid is associated with the correct tRNA
what are ribosomes made of?
-rRNA and protein
-each ribosome is made of two subunits, a large subunit and a small subunit
-each subunit contains one or more rRNA molecules
-the rRNA molecules are ribozymes and carry out the main functions of the ribosomes
-the proteins support the function of the ribozymes
ribosomes
-are essentially large ribozymes
-because there are so many ribosomes in a cell, rRNA is the most abundant type of RNA in the cell
-also have entry sites for the mRNA and exit tunnels for the polypeptide
P site, A site, and E site
the 3 locations that ribosomes contain for tRNAs to bind to an mRNA
P site
holds the tRNA that is attached to the growing polypeptide
A site
holds the tRNA attached to the next amino acid to be added to the polypeptide
E site
the exit site for the prior tRNA that has incorporated its amino acid already
3 phases of translation
- initiation
- elongation
- termination
(like transcription)
what 3 things are brought together during translation initiation?
a small ribsomal subunit, an mRNA, and an initiator tRNA carrying Met
translation initiation in bacteria vs eukaryotes
-in bacteria, the small ribosomal subunit can bind the mRNA and tRNA in any order
-in eukaryotes, the small ribosomal subunit binds the 5’ cap of the mRNA and then scans along the mRNA until it finds the start codon (AUG). then the initiator tRNA hydrogen bonds to the start codon
translation initiation complex
-once the mRNA and tRNA are in place on the small ribosomal subunit, the large ribosomal subunit binds to create the translation initiation complex
-this step requires an input of energy, which comes from the hydrolysis of GTP (very similar to ATP)
-initiation factors help to assemble the complex
-the initiator tRNA sits in the P site of the ribosome
initiation factors
other proteins that help to assemble the translation initiation complex
elongation factors
other proteins necessary for elongation and helps elongation to occur
translation elongation
-the codon in the A site of the ribosome base pairs with the appropriate tRNA (this step requires an energy input, GTP hydrolysis, which ensures the accuracy and efficiency of codon recognition)
-the energy input (GTP hydrolysis) moves the mRNA through the ribosome
-a peptide bond forms between the carboxyl group on one amino acid (in the P site) and the amino group on the next amino acid (in the A site)
-the tRNA in the A site moves to the P site moves to the E site and is released
-empty A site to receive next tRNA
peptide bond
-a covalent bond that forms between the two adjacent amino acids and the growing polypeptide is attached to the tRNA in the A site
-rRNA molecules in the ribosome catalyze the peptide bond
N-terminus and C-terminus
-the Met amino acid is at the N-terminus of the polypeptide
-the last amino acid added to the polypeptide chain is at the C-terminus
-polypeptides are always listed in order from the N-terminus to the C-terminus
-the codons for the amino acids at the N-terminus of a polypeptide are found at the 5’ end of a molecule
release factor
-binds to the codon when a stop codon enters the A site of the ribosome
-is a protein, shaped like a tRNA
-promotes the hydrolysis of the covalent bond between the amino acid and the tRNA sitting in the P site of the ribosome, freeing the polypeptide