Transcription and Translation
transcription and translation
two stages of gene expression are called
recap of mutations
-silent mutations: lead to translation of same amino acid, no nothing changes the final protein -missense mutations: lead to different amino acid being used, with effects depending on which amino acid was substituted or where it was substituted -nonsense mutations: lead to shorted protein because of premature stop codon -frameshift mutations: complete chaos
differences between DNA replication and RNA transcription
1.) DNA polymerase is extremely accurate and requires helicase and a primer to get started. RNA polymerase is less accurate (because its work isn't a permanent copy), but requires no primers to get started and unzips DNA itself - the ultimate self-starter. 2.) During DNA replication, both strands are complemented. During transcription, only the template strand is complemented. RNA polymerase ignoring the coding strand. 3.) During DNA replication, the DNA strand stays attached to the old strand and is complemented. In transcription, the new RNA strand is immediately removed and the two DNA strands come back together. 4.) DNA replicated using DNA nucleotides (including T), while RNA transcription uses RNA nucleotides (including U).
start codon
AUG not only coding for the amino acid Met anywhere in the protein, but also signaling the start codon for translation; so all proteins start with amino acid Met, regardless of what comes after
3 keys sites ribosomes performing translation
P site - holds the tRNA carrying the growing protein chain A site - holds the tRNA carrying the next amino acid to be added to the chain E site - where used tRNAs that have donated their Aino acid exit the ribosome
TATA Box and TATA-binding protein
a DNA sequence in eukaryotic promoters critical in forming the transcription initiation complex
stop signals for translation
UAA, UAG, UGA
GTP
acts just like ATP but instead of using an adenine nucleotide, it uses a guanine
synthesizing of complimentary mRNA
after the making complimentary mRNA molecule, it is then synthesized in an antiparallel direction to the template strand of DNA using the 5'-3' direction
replication proceeds in a 5'-3' direction, while transcription proceeds in a 3'-5' direction
all of the following are differences between RNA replication and transcription EXCEPT
-rRNA -mRNA -small subunit -E-site -P-site
all of the following are parts of the ribosome except:
functions of introns
allow for single gene to encode more than one version for a protein, depending one which segments are treated as exons during RNA processing, can get multiple protein products - alternative RNA splicing; so common that the average of three differently spliced mRNA's coming from each gene in human genome
27,000 nucleotides, but only takes 1,200 nucleotides to code for the aver-sized person
average length of a transcription unit along a human DNA molecule
each tRNA molecule
bears a specific amino acid at one end; other end is a nucleotide triplet - anticodon, which base-pairs code with a complementary codon in the mRNA
initiation in bacteria
binding of ribosomal subunit with mRNA and initiator tRNA occurs at a unique RNA sequences, just upstream of AUG; this unique RNA sequence establishes the codon reading frame in bacteria
multiple different tRNAs
brings amino acids to a ribosome and mRNA to see which of the 20 different amino acids comes next int he protein; ribosome will manage the process and link together the amino acids but will not be reading the mRNA to see which nucleotide comes next like tRNA
initiation
brings together an mRNA, a tRNA with the first amino acid, and two ribosomal subunits
messenger RNA
carrier of information form DNA to the cell's protein-synthesizing machinery that is transcribed from the template strand of a gene
sickle cell disease
caused by a mutation of a single nucleotide pair in the gene that encodes part of the protein hemoglobin; hemoglobin is critical to red blood cells; change in a single nucleotide in DNA's template strand leads to abnormal protein, causing the disease
mutations
changes in the DNA of a cell (or virus)
top strand is the coding strand
consider the following double-stranded DNA sequence that codes for a short polypeptide; which strand is the coding strand?
ribosome
consists of a large and small subunit, each made of proteins and ribosomal RNA (rRNA; in both bacteria and eukaryotic cells large and small subunits join together to form a functional ribosome
structure of tRNA molecule
consists of a single strand of about 80 nucleotides folded back on itself to form a three-dimensional structure; includes a loop containing the anticodon and an attachment site at the 3' end for an amino acid (anticodon nucleotides bind to the codon nucleotides during translation)
downstream or upstream
direction of transcription used by molecular biologists
DNA (genotype)
donates the synthesis of functional RNAs and proteins which are the link between genotype and phenotype
template strand
during transcription, the DNA strand that provides a template for ordering the sequences of nucleotide bases in an mRNA transcript
transcription
during which a DNA strand provides a template for the synthesis of each new complementary RNA strand; a protein-coding gene produces a messenger RNA (mRNA) molecule
codon
each set of three-letters
redundancy but no ambiguity
exists in genetic code but no ambiguity; several codons may specify same amino acids (redundancy) but no condone specifies more than one amino acid (no ambiguity)
process of initiation
first, small ribosomal subunit binds with mRNA and a special initiator tRNA, which carries methionine
genetic instructions in DNA
for a polypeptide chain where letters are written as a series of three-nucleotide "words"; every combination of three letters (AAA, ATA, CAC, etc) matches an amino acid
A, T, G, C
forms the information in genes, AKA: genotype
poly-A tails and 5' cap functions
functions of modifications: they signal that the mRNA is ready to leave the nucleus, they help the ribosomes attach to the mRNA, they crate "off switches" in the mRNA that can be removed later to stop translation
reason for a stop codon
in order for ribosome to so hold onto an mRNA, it needs to be longer than just the codons that is wanted (on both sides!); not only does it need to be told to stop before it gets to the end, but it also needs to be told where to start so AUG serves a dual role
transcription factors
in order to start transcription, what binds to the promotor segment on DNA?
phenotype
information inherited by an organism as traits
stages of translation
initiation, elongation, and termination - all three require protein "factors" that aid in the translation process; both initiation and elongation require energy provided by a relative ATP called GTP
insertion and deletion mutations
instead of swapping nucleotides, these add or subtract; likely to result in a shift in three-letter pattern of codons; unless insertion/deletion is a multiple of three letters, it causes a frameshift mutation; all nucleotides downstream of deletion/insertion will be improperly grouped into codons; results in extensive missence, ending sooner or later in sequence - premature termination
transfer RNA (tRNA)
interpreter between two languages of nucleotides to amino acids during translation
bacteria
lacks a nuclei so their DNA is not segregated from ribosomes and other protein-synthesizing equipment, allowing the coupling of transcription and translation; ribosomes attach to the leading end of an mRNA molecule while transcription is still in place
ribosomes
large enzymes that organize the assembly of amino acids into polypeptide chains and are sites of translation
pre-mRNA
made by eukaryotes after transcription is over; cannot be translated or exported until enzymes in the eukaryotic nucleus make three modifications: the 5' cap, the 3' poly-A tail and iron splicing; these modifications help form final mRNA molecule that is ready for export from the nucleus and translation
complimentary mRNA molecule
made using the regular-base pairing rules, just like when making new DNA (except that uracil is the complimentary base to adenine)
RNA polymerase II
makes mRNA for protein synthesis
triplet code
match between DNA letters and amino acids
determines the protein code
match between tRNAs and the amino acids they carry
mutations in gametes
may be transmitted to future generations; adverse effect on phenotype of organism causes mutation conditions know as genetic disorder or hereditary disease (ie. sickle cell disease)
single mRNA
may be used to make copies of a polypeptide simultaneously as multiple ribosomes, polyribosomes, or polysomes trail along same mRNA; polyribosomes can be found in both bacteria and eukaryotes
central dogma of molecular biology
molecular chain of command in a cell that has a directional flow of genetic information: DNA > RNA > Protein
reason for so many nucleotides in a transcription unit
most eukaryotic genes and their RNA transcripts have long non-coding stretches of nucleotides, called intervening regions or introns; remaining regions are called exons and are eventually expressed by being translated into amino acid sequences (introns are in the way, exons are expressed)
inhibition in detail
most tightly controlled part of RNA transcription; a lot of DNA in a cell and most isn't where RNA polymerase is starting the transcription so promotors and transcription factors are there to tell RNA polymerase where to start; transcription factors bind to DNA in promoter then after attachment, RNA polymerase II binds to it, making TATA box; RNA moves along DNA, unzips double helix, 10-20 nucleotides at a time; enzyme adds nucleotides to 3'-5' end of RNA growing strand; behind point of RNA synthesis, double helix re-forms and RNA molecule peels away
silent mutation
mutation that causes a change in a nucleotide pair may transform one codon into another that is translated into the same amino acid
gametes (sperm and egg cells)
mutations can be passed on to the next generation (offspring), if they occur in
missense mutations
mutations that alter the codon enough to change the amino acid; some cases are minimal; if amino acids switched properties or if mutation occurs in a region where the exact sequence is not essential for function, the change might not even cause a phenotype; BUT if occurs at critical sites, esp. active sites on an enzyme, can have major effects
nonsense mutations
mutations that change an amino acid codon into a stop codon, causing premature termination of translation and nearly always leading to nonfunctional protein; many recessive genes are actually nonsense mutations that produce no functional protein (ie white phenotype of purple/white pea flowers)
RNA polymerase; promoter
name of the enzyme that transcribes DNA into RNA is called _____ and it starts the site of DNA called the _____
redundancy
never random; codons that are synonyms for a particular amino acid differ only in the third nucleotide of the triplet; ie all four option for CU_ code for Leu; third letter doesn't even really matter in this case
4^3 = 64
number of every possible combination of the four letters to make a three-letter words gets a maximin combination
primary transcript
occurs in eukaryotic cells as the initial RNA transcript of any gene
one amino acids
one codon matches
alternative mRNA splicing
one explanation for how a single gene can code for more than one version of a protein is
in eukaryotes, ribosomes only attach to mRNA AFTER the mRNA is completely made, while in prokaryotes, ribosomes attach to the mRNA BEFORE the mRNA is completely made
one important difference between green expression in eukaryotes and bacteria is
substitution mutation
one of multiple point mutations where the replacement of one nucleotide and its partner with another pair of nucleotides; some nucleotide-pair substitutions have no effect on protein function - due to redundancy of third letter in many codons
UTRs
parts of the mRNA that will not be translated into protein (untranslated regions)
gene expression
process by which DNA directs protein synthesis, including two stages of transcription and translation
translation
process where a cell converts a genetic message (nucleotides) into a protein (amino acids)
genes
program protein synthesis via genetic messages in the form of messenger RNA
transcription factors
proteins that bind to DNA sequences in the promoter
transcription process
provides a template for assembling a sequences of RNA molecules, just like a DNA strand providing a template for the synthesis of each new complementary strand during DNA replication
ribosomes
responsibilities include aligning the tRNAs with the mRNA message and acts as an enzyme that binds the new amino acid to the growing protein chain (forms the peptide bond between amino acids)
elongation stage
second step in translation where amino acids are added one by one to the previous amino acid at the end of the growing chain; after each amino acid is added, the mRNA is moved through the ribosome in a 5'-3' direction by 3 letters, or one codon, creating new space for tRNAs to try to complement the codon; the empty tRNAs that are released from E site return to cytoplasm where they're reloaded with appropriate amino acid
RNA polymerase
separates the DNA strand at the appropriate point and joins RNA nucleotides complementary to the DNA template strand; can assemble a strand only in 5'-3' direction like DNA but unlike DNA polymerase, can start a chain from scratch - do NOT need helicase or a primer
tempalate sequence of DNA
sequence of nucleotides in mRNA is complimentary to
RNA
similar to DNA, except the tis contains ribose instead of deoxyribose as its sugar and substitutes the nitrogenous base uracil for thymine (DNA) - consists of a single strand
introns are removed
splicing is when
inhibition
stage where RNA polymerase binds to precise location facing downstream on the promoter, which determines where transcription starts and which of the two strand of the DNA helix is the template
termination
stage where RNA polymerase continue until it reaches sequences that mark the end of a message; in prokaryotes, this is a completed RNA transcript since there is no nucleus, so ribosomes are already working on translation before termination of transcription even happens; in eukaryotes, RMA undergoes some processing steps before it's exported form nucleus to cytoplasm to be translated into proteins on ribosomes
elongation
stage where the RNA polymerase unzips the DNA, reading the DNA in the 3'-5' direction and synthesizing RNA in the 5'-3' direction; RNA polymerase knows which nucleotides to place in new RNA strand, like DNA polymerase, tries them at random until finds a complimentary pair
initiation, elongation, and termination
stages of transcription of the RNA chain
5' cap
the 5' end of the pre-mRNA where a modified form of the guanine nucleotide (G) is added; not to be translated
poly-A tail
the end of 3' end where an enzyme adds 50 to 250 adenine nucleotides (A); not to be translated
missene
the original mRNA sequence is AUG, CCA, GGG, UGU, UGA but after mutation the sequence of the mRNA is AUG, CGA, UGU, UGA; what type of mutation?
RNA splicing
the removal of a large portion of the RNA molecule in a cut-and-paste job
initiation in eukaryotes
the small subunit with the initiator already bound, binds to the 5' cap of the mRNA and then moves, or scans, downstream until it reaches the first AUG start codon AUG; this first AUG establishes the codon reading frame in eukaryotes; the union of mRNA, initiator tRNA, and a small ribosomal subunit is followed by the attachment of a large ribosomal subunit
translation
the synthesis of a protein, using information in mRNA; during which, there is a change of coding language from nucleotides (DNA and RNA) to amino acids (protein)
other strand of DNA
these codons on the mRNA are identical to the letters in the non-template strand of DNA (except have U instead of T), so called coding strand; even though not being used at all during transcription, it predicts exactly how the mRNA will read
termination
third stage that occurs when one of the three stop codons reaches the A site of the ribosome; a release factor binds to the stop codon and causes hydrolysis of the bond between polypeptide and its tRNA in the P site; this frees the polypeptide, which is released through the exit tunnel of the ribosome's large subunit; translation complex then disassembles
initiation, elongation, termination
three major steps of translation are:
stop codons
three of the total number of possible codons in the mRNA specify for
eukaryotic cell
transcription occurs in the nucleus and the transcription of a protein-coding eukaryotic gene results in pre-mRNA; the primary-transcript is then further processed resulting in finished mRNA, which is then exported to the cytoplasm, binding to a ribosome and translated into protein
tRNA
translator because it reads a nucleic acid work (the mRNA codon). and interprets it as a protein word (the amino acid); works as codon by codon the genetic message os translated as it deposits amino acids in the order prescribed and the ribosome connects to amino acids into chain
mutations
ultimate source of new genes; include large-scale mutations, in which long segments of DNA are affected (ie. translocations, duplications, and inversions), as well as point mutation, changes in just one nucleotide pair of a gene
codon triplet of genetic code
use the codon triplet to figure out amino acid; first nucleotide is chosen from the right side, second nucleotide is chosen from the top panel and third nucleotide is chosen from right side
mRNA codons
what molecular "language molecular" does tRNA interpret into amino acids?
silent mutation
when a mutation does not cause a change in the amino acid coded for it, it is called a
process of translation
when the sequence of codons along an mRNA molecule that is translated into a sequence of amino acids that make up the polypeptide chain, reading in a 5'-3' direction; codon then specifies which of 30 amino acids will be incorporated at the corresponding position along a polypeptide chain
promoter
where RNA polymerase attaches and initiates transcription along specific sequences of nucleotides of the DNA strand where gene expression begins and ends
exons
which of the following in mRNA is coded into protein (amino acids)?
-the genetic code can code more than one amino acid -the genetic code is unambiguous -the genetic code is triplet-based -each codon represents a different amino acid
which of the following is not a true statement regarding genetic code?
RNA uses ribose as a sugar, while DNA uses deoxyribose
which of the following is the major difference between DNA and mRNA